Search

Background: High-quality data are of prime importance in any health survey because survey data are considered as a gold standard for nationally representative data. The quality of data collection largely depends on the design of the questionnaire, training, and skills of the interviewer. Objectives: In the present study, we tried to evaluate three key components, such as questionnaire design, human resource and training of the field staff for Integrated Biological and Behavioural Surveillance carried out among the HIV high-risk subpopulation. Methods: A mixed-methods approach was used. Qualitative and quantitative data collection was carried out in the year 2015 with cross-sectional survey design in western states of India. The in-depth interviews of 10 stakeholders, structured interviews of the survey respondents (n = 560), and field investigators (n = 71) were conducted. Data triangulation was used to find out the concurrence of the qualitative and quantitative data. Results: Comprehensive and standardized survey questionnaire, structured training agenda, and strategic preparation for recruiting human resources were the overall strengths of the survey. However, during the implementation of the survey, there were some difficulties reported in data collection process. Overall, the respondents and investigators felt that the questionnaire was long and exhaustive. Difficulties were faced while collecting data on sexual history. The field staffs were not adequately experienced to work with sensitive population. Conclusions: In order to have accurate, reliable data, especially on sexual behavior; emphasis should be given on simple questionnaire with the use of community-friendly language, skilled and experienced interviewers for data collection, and extensive field training.

Purpose. Emergence of multidrug resistance in Neisseria gonorrhoeae, an STI of public health significance is the biggest challenge to gonorrhoea control. Monitoring for antimicrobial resistance is essential for the early detection of emergent drug resistance patterns. Methodology. One hundred and twenty four N. gonorrhoeae strains were isolated between September 2013-August 2016 [82-New Delhi, 3-Pune, 3-Mumbai, 20-Secunderabad and 16-Hyderabad] to determine antimicrobial susceptibility and to compare the CLSI disc diffusion method with Etest for these strains. The results of the two methods were compared by using kappa statistics. Results. Ninety eight percent [CI: 96.2–100] of isolates were resistant to ciprofloxacin, 52 % [CI: 43.2–60.8] to penicillin, 56 % [CI: 47.2–64.7] to tetracycline and 5 % [CI: 1.2–8.8] to azithromycin. All the strains were susceptible to spectinomycin, ceftriaxone and cefixime except for two strains which showed decreased susceptibility to ceftriaxone and cefixime. Kappa scores for penicillin, azithromycin, ciprofloxacin, ceftriaxone and cefixime showed that the CLSI method had high agreement with Etest while tetracycline had substantial agreement. Conclusion. Our data suggest that the disc diffusion method which is both cost effective and more feasible, can effectively be used routinely for monitoring antibiotic susceptibility in N. gonorrhoeae, in limited resource countries like India. We demonstrate the emergence of decreased susceptibility to ceftriaxone and cefixime and threshold levels of resistance to azithromycin in India. This underscores the importance of maintaining continued surveillance for antibiotic resistance in N. gonorrhoeae and a potential requirement for strategic change in guidelines in the not so distant future.

Background Several studies in recent years have documented the genotype-specific prevalence of HPV infection and wide diversity and multiplicity of HPV genotypes among HIV-seropositive women. Yet, information on changes in HPV genotype-specific incidence and clearance rates over time, and their correlation with clinical or immunologic factors among HIV-seropositive women is scarce. Objectives We conducted a prospective study to investigate the incidence and clearance rates of cervical HPV genotypes among HIV-seropositive women in India and expand the evidence base in this area of research. Study design Cervical samples were collected from n = 215 HIV-seropositive women in Pune, India who underwent two screening visits separated by a median of 11-months (interquartile range: 8–18 months). HPV genotypes were determined by Roche Linear Array HPV assay. Individual genotype-specific and carcinogenicity-grouping-specific HPV incidence and clearance rates were calculated and the associations between incidence/clearance and age and HIV-related metrics were explored. Results Incidence and clearance rates for ‘any HPV’ and ‘carcinogenic HPV’ genotypes were 11.1 and 18.3, and 6.7 and 33.8, per 100 person-years, respectively. Incidence and clearance rates for HPV genotypes of alpha-9 species (HPV16, HPV31, HPV33, HPV35, HPV52 and HPV58) and alpha-7 species (HPV18, HPV39, HPV45, HPV59 and HPV68) were 5.8 and 2.04, and 32.1 and 53.5, per 100 person-years, respectively. Clearance of any HPV type was associated with increasing age of participants (odds ratio: 1.08, 95%CI: 1.004–1.17), although the association marginally lost its statistical significance when adjusted for CD4 counts and antiretroviral therapy status. Conclusions Genotype-specific clearance rates of HPV were higher than corresponding incidence rates. The suggestion of a positive associations of increasing age with HPV clearance points to the need for etiologic studies on age-related hormonal changes on clearance of cervical HPV infection.

Under India’s National AIDS Control Program (NACP), WHO recommended strategy of using three rapid tests is adopted for diagnosis of HIV in an asymptomatic individual. Since the NACP has a stringent kit evaluation procedure and due to the availability of newer third generation Rapid Diagnostic Tests (RDTs) it may be possible to adapt two test strategy for HIV diagnosis instead of the recommended three test strategy. The authors reviewed programmatic data on HIV testing to explore whether use of two rapid tests for HIV diagnosis as against three tests could be a feasible approach without compromising the quality and readability of testing. Data on the HIV diagnosis performed on serum specimens collected for the period of one year (2011-12) from 82 Integrated Testing and Counselling Centers (ICTCs) or Prevention of Parent to Child Transmission Centers (PPTCTs) associated with State reference Laboratories (SRLs) was analysed. Out of 654258 results that were analysed 25168 (3.84%) specimens were positives. It was observed that both two and three test algorithms provided similar results for majority (>99%) of the HIV positive specimens. Discordance was observed in labelling of specimen with inconclusive status (N = 21), however the true status of these samples could not be obtained. The analysis indicated that the use of two test algorithm will have programmatic benefits in terms of reduced financial burden to the programme and ease of procurement, shipment and storage before and after distribution without comprising the quality of the testing. The prospective study would confirm this observation.

In India, the roll out of the free antiretroviral therapy (ART) program completed a decade of its initiation in 2014. The success of first-line ART is influenced by prevalence of HIV pretreatment drug resistance (PDR) in the population. In this cross-sectional study, we sought to determine the prevalence of PDR among adults attending the state-sponsored free ART clinic in Pune in western India. Fifty-two individuals eligible for ART as per national guidelines with median CD4 cell count of 253 cells/mm(3) (inter quartile range: 149-326) were recruited between January 2014 and April 2015. Population-based sequencing of partial pol gene sequences from plasma specimen revealed predominant HIV-1 subtype C infection (96.15%) and presence of single-drug resistance mutations against non-nucleoside reverse transcriptase inhibitor in two sequences. The study supports the need for periodic surveillance, when offering PDR testing at individual level is not feasible.

Using new biomarker data from the 2010 pilot round of the Longitudinal Aging Study in India (LASI), we investigate education, gender, and state-level disparities in health. We find that hemoglobin level, a marker for anemia, is lower for respondents with no schooling (0.7 g/dL less in the adjusted model) compared to those with some formal education and is also lower for females than for males (2.0 g/dL less in the adjusted model). In addition, we find that about one third of respondents in our sample aged 45 or older have high C-reaction protein (CRP) levels (>3 mg/L), an indicator of inflammation and a risk factor for cardiovascular disease. We find no evidence of educational or gender differences in CRP, but there are significant state-level disparities, with Kerala residents exhibiting the lowest CRP levels (a mean of 1.96 mg/L compared to 3.28 mg/L in Rajasthan, the state with the highest CRP). We use the Blinder–Oaxaca decomposition approach to explain group-level differences, and find that state-level disparities in CRP are mainly due to heterogeneity in the association of the observed characteristics of respondents with CRP, rather than differences in the distribution of endowments across the sampled state populations.

Introduction The HIV sentinel surveillance [HSS] conducted in 2010–11 among female sex workers [FSW] in the state of Maharashtra, India provided an opportunity to assess characteristics of different types of FSWs and their HIV risk. It is important for India’s National AIDS Control Program, to understand the differences in vulnerability among these FSW, in order to define more specific and effective risk reduction intervention strategies. Therefore, we analyzed data from HSS with the objective of understanding the HIV vulnerability among different types of FSW in Maharashtra. Material and methods Cross sectional data collected as a part of HSS among FSWs in year 2010–11 from 21 sentinel sites in the state of Maharashtra were analyzed to understand the vulnerability and characteristics of different types of female sex workers based on their place of solicitation using multinomial logistic regression. Results While the HIV prevalence was 6.6% among all FSWs, it was 9.9% among brothel based [BB], 9% among street based [SB] and 3.1% and 3.7% among home based [HB], and bar based [Bar-B] sex workers respectively. SB FSWs were least likely to be located in HIV low burden districts [ANC] [ARRR: 0.61[95% CI: 0.49, 0.77]], but were 6 times more likely to be recently [1%). We identify them as a group to be focused on for prevention interventions and it is likely that they would be easily amenable to novel interventions due to their higher literacy rate as compared to other typologies.

BACKGROUND Although HIV incidence has declined in India, men and transgender women who have sex with men (MSM) continue to have high rates of HIV and sexually transmitted disease (STD). Indian MSM face substantial pressures to marry and have families, but the HIV/STD burden among married Indian MSM is not well characterized. METHODS A diverse sample of Indian MSM were recruited through respondent-driven sampling. Independent variables that produced a P value of 0.10 or less were then added to a multivariable logistic regression model. RESULTS Most of the 307 MSM (95 married and 212 unmarried) recruited into the study were younger than 30 years, and less than one-third had more than a high school education. Almost two-thirds of the married men had children, compared with 1.4% of the unmarried men (P < 0.001). The numbers of condomless anal sex acts did not differ by marriage status. Although unmarried MSM more often identified themselves as "kothi" (receptive role), their rates of HIV or bacterial STD were similar to married MSM, with 14.3% being HIV infected. The respondent-driven sampling-adjusted prevalence of any bacterial STD was 18.3% for married MSM and 20% for unmarried MSM (not significant). Participants reported high levels of psychological distress, with 27.4% of married and 20.1% of unmarried MSM reporting depressive symptoms (not significant). CONCLUSIONS Men who have sex with men in Mumbai had high rates of HIV, STD, and behavioral health concerns. Clinicians need to become more comfortable in eliciting sexual histories so that they can identify MSM who need HIV/STD treatment and/or prevention services.

There has been a reduction in the most severe cases of HIV-associated neurocognitive disorders (HAND) with advances in antiretroviral treatment (ART). But the prevalence of milder forms of HAND still remains high. Data from systematically conducted studies on the effects of ART on cognition are scanty in India, where HIV-1 clade C is prevalent. The purpose of the present study was to assess the effect of antiretroviral therapy in HIV-seropositive (HIV+) individuals (n = 92) with CD4 cell counts

Saliva plays an important role in maintaining microbial homeostasis in the oral cavity, while salivary gland hypofunction predisposes the oral mucosa to pathologic alteration and increases the risk for oral candidiasis. This study sought to determine the salivary flow rate (SFR) and secretory immunoglobulin A (SIgA) levels in HIV-positive and HIV-negative individuals and evaluate their relationship with the determinants of oral candidiasis. Sixty HIV-positive (30 with and 30 without oral candidiasis) and 30 healthy HIV-negative individuals were enrolled. Cotton pellet was weighed pre- and post-saliva collection for the assessment of SFR, while SIgA levels were estimated by commercial ELISA (Diametra, Italy) kit. The mean ± SD, SFR and SIgA levels in HIV-positive individuals with candidiasis, without candidiasis and HIV-negative controls were 0.396 ± 0.290, 0.546 ± 0.355 and 0.534 ± 0.214 ml/min and 115.891 ± 37.621, 136.024 ± 51.075 and 149.418 ± 31.765 µg/ml, respectively. A positive correlation between low CD4 counts (indicator of immunodeficiency) and SIgA was observed in HIV-positive individuals with candidiasis (r = 0.373, p = 0.045). We also report here for the first time the significant decrease in SFR and SIgA levels in individuals presenting with pseudomembranous type of oral candidiasis and Candida albicans infection.

Background and Objectives: Emergence of multi-drug resistant Neisseria gonorrhoeae resulting from new genetic mutation is a serious threat in controlling gonorrhea. This study was undertaken to identify and characterise mutations in the mtrR genes in N.gonorrhoeae isolates resistant to six different antibiotics in the quinolone group. Materials and Methods: The Minimum inhibitory concentrations (MIC) of five quinolones for 64 N.gonorrhoeae isolates isolated during Jan 2007-Jun 2009 were determined by E-test method. Mutations in MtrR loci were examined by deoxyribonucleic acid (DNA) sequencing. Results: The proportion of N.gonorrhoeae strains resistant to anti-microbials was 98.4% for norfloxacin and ofloxacin, 96.8% for enoxacin and ciprofloxacin, 95.3% for lomefloxacin. Thirty-one (48.4%) strains showed mutation (single/multiple) in mtrR gene. Ten different mutations were observed and Gly-45 → Asp, Tyr-105 → His being the most common observed mutation. Conclusion: This is the first report from India on quinolone resistance mutations in MtrRCDE efflux system in N.gonorrhoeae. In conclusion, the high level of resistance to quinolone and single or multiple mutations in mtrR gene could limit the drug choices for gonorrhoea.

Introduction Cytomegalovirus (CMV) pneumonia is one of the frequent viral pneumonia reported in persons with HIV infection. Knowledge of pulmonary CMV infection is important for deciding appropriate diagnostic strategies. However, there is scanty literature addressing the role of CMV aetiology among HIV positive individuals presenting with Community Acquired Pneumonia (CAP) using Bronchoalveolar Lavage (BAL) samples from India. Aim To detect CMV in BAL fluid from HIV-positive individuals presenting with CAP. Materials and methods This cross-sectional study was conducted using 107 archival BAL samples collected from consecutive HIV-positive patients presenting with CAP as per the Indian Chest Society and National College of Chest Physicians guidelines at the Department of Chest and Tuberculosis, Sassoon General Hospitals, Pune, India. The samples were tested for CMV by Polymerase Chain Reaction (PCR) targeting the IRL11 region at the National AIDS Research Institute, Pune. Results Of the 107 BAL samples tested, 8 (7.4 %) were positive for CMV, while CMV was the sole pathogen in 5 (4.7%) cases. Co-infection with other pathogens was seen in 3 patients and Mycobacterium tuberculosis, Pneumocystis jiroveci and Streptococcus pneumoniae were the co-pathogens. Five patients had fatal clinical outcome of which three had CMV as the sole pathogen. Conclusion Ours is the first study to detect Cytomegalovirus (CMV) in bronchoalveolar lavage samples from HIV-positive individuals presenting with community acquired pneumonia from India and indicates the need for further multicentre studies to understand pulmonary CMV infection, which will eventually help in designing appropriate diagnostic strategies and therapeutic interventions.

To detect CMV in BAL fluid from HIV-positive individuals presenting with CAP.This cross-sectional study was conducted using 107 archival BAL samples collected from consecutive HIV-positive patients presenting with CAP as per the Indian Chest Society and National College of Chest Physicians guidelines at the Department of Chest and Tuberculosis, Sassoon General Hospitals, Pune, India. The samples were tested for CMV by Polymerase Chain Reaction (PCR) targeting the IRL11 region at the National AIDS Research Institute, Pune.Of the 107 BAL samples tested, 8 (7.4 %) were positive for CMV, while CMV was the sole pathogen in 5 (4.7%) cases. Co-infection with other pathogens was seen in 3 patients andOurs is the first study to detect

Comprehensive management of sexually transmitted infections (STIs) using vaginal or rectal microbicide-based intervention is one of the strategies for prevention of HIV infection. Herbal products have been used for treating STIs traditionally. Herein, we present in vitro activity of 10 plant extracts and their 34 fractions against three sexually transmitted/reproductive tract pathogens – Neisseria gonorrhoeae, Haemophilus ducreyi and Candida albicans. The plant parts were selected; the extracts/fractions were prepared and screened by disc diffusion method. The minimum inhibitory and minimum cidal concentrations were determined. The qualitative phytochemical analysis of selected extracts/fractions showing activity was performed. Of the extracts/fractions tested, three inhibited C. albicans, ten inhibited N. gonorrhoeae and five inhibited H. ducreyi growth. Our study demonstrated that Terminalia paniculata Roth. extracts/fractions inhibited growth of all three organisms. The ethyl acetate fraction of Syzygium cumini Linn. and Bridelia retusa (L.) Spreng. extracts was found to inhibit N. gonorrhoeae at lowest concentrations.

Sir, The identification of Neisseria gonorrhoeae by sensitive, precise, and rapid technique is important for the correct diagnosis and appropriate treatment of patients with gonorrhoea. For decades, the standard method of identification of the pathogen has been acid production from cystine tryptic agar (CTA) medium containing carbohydrates1. For a more rapid identification of this pathogen a variety of immunological and biochemical commercial kits have now become available. An ideal test should have high sensitivity and specificity, should be easier to perform and cheap. An attempt was made in this study to identify such a test by comparing four available tests on 71 N. gonorrhoeae isolates. The three most widely used immunological kits are the MicroTrak N. gonorrhoeae culture confirmation test (an immunofluoresence test; Trinity Biotech PIC, Ireland), the Phadebact Monoclonal GC test (a coagglutination test; MKL Diagnostics AB, Kung Hans Vag, Sollentuna, Sweden) and GonoGen II (a membrane immunoassay; New Horizons Diagnostics, USA), all of which employ monoclonal antibodies (mAbs) developed against specific epitopes on the two types of the major outer membrane protein, protein I A and B (PIA and PIB)2. Interpretation of the MicroTrak test is less subjective but requires a fluorescence light microscope whereas Phadebact monoclonal test is easier to perform and can be used by most small laboratories2,3. GonoGen II is a colorimetric test and does not require heat treatment step of the coagglutination test. This test eliminates the reading of agglutination; instead, the mixture of antigen-antibody complex is passed through a filtration unit. The appearance of a red dot on the filter indicates a positive reaction2. Though the specificity of these kits for detection of N. gonorrhoeae is high, both false-positive (cross-reactions with other Neisseria species such as N. meningitidis, N. lactamica, N. cinerea and Kingella denitrificans) and false-negative results have also been reported4. The biochemical kits widely used are the Neisseria Preformed Enzyme Test (PET), Gonochek II, RapID NH, Neisstrip, API-NH and Rapid carbohydrate utilization test (RCUT)3,4,5. These tests are based on the presence of preformed chromogenic enzyme in the culture, and thus require a heavy inoculum of the organism grown on selective medium to permit rapid speciation of isolates. The enzymes that are detected by these systems include beta-galactosidase, gamma-glutamylaminopeptidase and prolyl-hydroxyprolyl aminopeptidase2. The Neisseria PET and Gonochek II are both single-use tubes containing chromogenic substrates that detect the presence of three preformed enzymes, each of which is indicative of a pathogenic Neisseria species, namely N. lactamica, N. meningitidis and N. gonorrhoeae. In Gonochek II, after incubation a red colour change (hydrolysis of L-proline 4-methoxynapthylamide) is observed2,3. The API-NH and RapID NH kits employ a battery of tests, combining carbohydrate utilization and preformed enzymes. These two tests detect the change in prolyl iminopeptidase (PIP) enzyme activity due to the mutation in the pip gene. Not all isolates of N. gonorrhoeae possess PIP activity6,7. In RCUT, preformed enzyme is measured by adding a suspension of the overnight growth of the suspect organism to a buffered (non-nutrient) solution containing the sugar to be tested and a pH indicator. A yellow colour change indicated a positive reaction and orange red colour as negative reaction8. As taxonomic differences between members of the Neisseria genus remain small, the identification of this pathogen can be problematic. Of the several commercial kits used worldwide for identification of N. gonorrhoeae, only four commercial kits, RCUT, API-NH, Gonochek II and Phadebact GC system were available in India. The purpose of this study was to compare the performance of these four commercial biochemical methods to identify pathogenic N. gonorrhoeae and to assess the cost effectiveness and utility in the clinical laboratory. A total of 71 Neisseria isolates were included in the study carried out at the National AIDS Research Institute (NARI), Pune, India, during January 2007 to June 2009. Of these, 64 isolates of N. gonorrhoeae were obtained from patients attending STD clinics of NARI, Pune (n=16), Safdarjang hospital, New Delhi (n=35) and 13 collected under STI-Operations Research project conducted at NARI (Mumbai-6, Hyderabad-6 and Nagpur-1). Patients attending STD clinics with complaints of urethral/cervical discharge were included in the study. Seven non gonococcal Neisseria strains, N. sicca (n=2), N. cinerea (n=2) and N. lactamica (n=3) recovered from clinical specimens from patients attending NARI clinics were also included. All the strains were tested by API-NH (Bio Merieux SA, France), Gonochek II (E-Y Laboratories, CA, USA), Phadebact monoclonal GC OMNI test (Boule Diagnostics AB, Sweden) according to manufacturers’ instructions, and by Rcut8. N. gonorrhoeae ATCC 49226 (β-lactamase negative) and WHO strains A, G H, E and O (received under WHO Gonococcal Antimicrobial Surveillance Programme from WHO GASP South East Asia Region Reference Laboratory, New Delhi) (β-lactamase positive) were used as quality control in each test run. The cost per test for each identification method was calculated considering the cost of media, reagents, ancillary supplies and time required to perform identification by each test method. All isolates of N. gonorrhoeae were correctly identified with RCUT system and Phadebact monoclonal GC OMNI system (Table). Gonochek II test correctly identified in 68 (95.3%) of N. gonorrhoeae isolates. The low sensitivity of this test has also been reported earlier. API-NH system correctly identified 51 (79.6%) of N. gonorrhoeae isolates. Thirteen (20.3%) N. gonorrhoeae isolates did not show any reaction for PIP enzyme in the API-NH system, which could be done to the lack of enzyme in these isolates. Specificity of these tests was evaluated based on their ability to give negative results on the seven non gonococcal strains. RCUT system correctly identified N. cinerea, N. sicca strains and one of the three N. lactamica strains as non gonococcal strains. Similar findings were observed by Young and Moyes9. API-NH system correctly identified both N. cinerea strains, one of the two N. sicca strains and one of the three N. lactamica strains as non gonococcal strains. Gonochek II test correctly identified one of the three N. lactamica strains as non gonococcal. Other studies have also reported similar findings in identifying N. gonorrhoeae, N. lactamica and N. sicca strains5,10. The Phadebact monoclonal GC OMNI system correctly identified both N. cinerea strains, two of the three N. lactamica strains and one of the two N. sicca strains. The sensitivity and specificity of Phadebact monoclonal GC OMNI system in our study was similar to that reported earlier5. Thus, RUCT appeared to be the most specific (6/7) followed by Phadebact OMNI system (5/7), and API-NH (4/7). Gonochek II test showed lowest specificity by identifying six of the 7 non gonococcal strains as N. gonorrhoeae. RCUT was found to be the cheapest among all tests (Table). Table Comparison of four test systems for the identification of N. gonorrhoeae isolates The Phadebact monoclonal GC OMNI system was rapid, simple to perform, accurate and provided results in 2-3 min. However, it showed moderate specificity and was more expensive than other methods. The colours generated in the Gonochek II system incubated at 30°C for 30 min were distinct, easy to read, stable and easy to handle in the clinical laboratory. However, it had low sensitivity (95.3%) in identification of N. gonorrhoeae. Additionally, it appeared to have low specificity since six of the seven non gonococcal strains were identified as gonococcal strains. API-NH system showed low sensitivity and specificity in identification of N. gonorrhoeae. This may be due to the lack of enzyme PIP in these isolates which have also been reported in other studies4,5,6,7. We found that the RCUT gave reliable results within 4 h, and identified 100 per cent of N.gonorrhoeae isolates as reported earlier by Yong and Moyes9. Additionally, it appeared to be highly specific since only one of seven non gonococcal strains was wrongly identified as gonococcal strain. Considering the performance, rapidity and cost, RCUT appeared to be the best among the four methods studied for confirmatory identification of N.gonorrhoeae. RCUT can be used routinely as it is cheaper and easily available but the reagents used for RCUT should be evaluated with the known control strain before being used routinely.

Objectives This study aims to validate a modified dried blood spot (DBS)-based glycosylated hemoglobin (HbA1c) assay protocol, after a pretest in India showed poor correlation between the original DBS-based protocol and venous results.

BACKGROUND Diarrhea in HIV infected individuals is a common complication seen in about 90% of patients in developing countries. The objective of this study was to identify enteric pathogens in HIV infected and HIV uninfected individuals in Pune. METHOD This study was conducted from January 2009 to May 2010 on 331 consecutive patients presenting with diarrhea admitted at Naidu Municipal Corporation Hospital, Pune and processed using conventional methods. RESULTS Intestinal parasitic pathogens were detected in 60% (39/65) of HIV infected and 14.3 (38/266) of HIV uninfected individuals. Bacterial pathogens were detected in 34% (22/65) of HIV infected individuals and 28.2% (75/266) of the HIV uninfected individuals. The common enteric pathogens detected in HIV infected individuals were Cystoisospora belli (28%, 18/65) followed by Cryprotosporidium parvum (12%, 8/65). In HIV uninfected individuals Entamoeba histolytica (7.1%, 19/266) followed by Shigella flexnari (4.9%, 13/266) were the most common pathogens. The difference in detection of enteric parasites in HIV infected individuals and HIV uninfected individuals was found to be significant (p < 0.01). CONCLUSIONS Intestinal parasitic pathogens are more common in HIV infected antiretroviral therapy naive patients. Early detection of enteric pathogens by routine examination of stool samples will help in the management and to improve the quality of life for HIV infected individuals.

BACKGROUND: The Revised National TB Control Program bases diagnosis of tuberculosis (TB) on sputum smear examination and response to a course of antibiotics, whereas World Health Organization recommends early chest radiography [chest x-ray (CXR)] for HIV-infected symptomatic patients. We evaluated the utility of initial CXR in the diagnostic algorithm for symptomatic HIV-infected patients with negative sputum smears. METHODS: HIV-infected ambulatory patients with cough or fever of ≥2 weeks and 3 sputum smears negative for acid-fast bacilli were enrolled in Chennai and Pune, India, between 2007 and 2009. After a CXR and 2 sputum cultures, a course of broad-spectrum antibiotics was given and patients were reviewed after 14 days. Sensitivity, specificity, positive and negative predictive values of symptoms, CXR, and various combinations for diagnosing pulmonary tuberculosis (PTB) were determined, using sputum culture as gold standard. RESULTS: Five hundred four patients (330 males; mean age: 35 years; median CD4: 175 cells per cubic millimeter) were enrolled. CXR had a sensitivity and specificity of 72% and 57%, respectively, with positive predictive value (PPV) of 21% and negative predictive value (NPV) of 93% to diagnose PTB. TB culture was positive in 49 of 235 patients (21%) with an abnormal initial CXR and 19 of 269 patients (7%) with a normal CXR (P

Purpose: Genital Ulcer Disease (GUD) is common sexually transmitted infection (STI). Multiple studies have shown that GUDs are strongly associated with the transmission and the acquisition of HIV infection. An accurate diagnosis of common etiology of GUD namely Herpes, syphilis and Chancroid is possible using Multiplex PCR (M-PCR). However, frequent presence of Polymerase Chain Reaction inhibitors in the ulcer swab specimen limits the performance of the assay. In order to overcome this problem, alternative specimen preparation method was used. Materials and Methods: To determine the common etiology, GUD specimens obtained under an STI operations research study were tested with M-PCR after the samples were prepared using Roche Amplicor specimen preparation kit. PCR inhibiting samples were identified from that, which showed negative results. These samples were subjected to phenol-chloroform extraction and ethanol precipitation before the conduct of M-PCR on them. Results: Of the 237 GUD specimens tested, in 145 etiologies could be detected, whereas 92 samples were found negative. Further spiking with one of the target DNA, 128 of the negative samples were found to contain the inhibitors. These 126 samples were then subjected to phenol chloroform extraction and ethanol precipitation followed by M-PCR. Using this method for sample preparation, etiology could be determined in 46 (23%) additional samples. This success rate of altered sample preparation method has been lower than that has reported. Conclusion: The results indicate that sample preparation using phenol chloroform extraction and ethanol precipitation, prior to M-PCR helps to eliminate the inhibitors and increase the yield of the assay. However, being a laborious procedure, it may be used for samples giving negative results after the screening by Roche Amplicor specimen preparation kit.

A dramatic increase in the number of quinolone-resistant Neisseria gonorrhoeae isolates in India and worldwide has been reported recently. This study was undertaken to identify and characterise mutations in the gyrA and parC genes of N. gonorrhoeae resistant to six different quinolone antibiotics. In total, 64 N. gonorrhoeae clinical isolates were obtained during 2007-2009 from patients attending sexually transmitted diseases clinics (New Delhi, 35; Pune, 16; Mumbai, 6; Hyderabad, 6; and Nagpur, 1). Antimicrobial susceptibility was determined by Etest and mutation patterns in gyrA and parC were determined by sequencing analysis. All strains showed varying resistance to different quinolone analogues, ranging from 17.2% (gatifloxacin) to 98.4% (ofloxacin and norfloxacin). Sequencing of gyrA and parC revealed that 100% of strains showed mutations in gyrA and 46.9% showed mutations both in gyrA and parC. All strains showed single or double mutations at Ser-91→Phe, Ser-91→Thr and Asp-95→Gly/Asn in gyrA and at Glu-91→Gly in parC. Asp-95→Asn mutation was the most prevalent in strains isolated from New Delhi, whilst Asp-95→Gly was prevalent in strains isolated from Pune. Strains were categorised into eight different mutation patterns. Resistant strains with high minimum inhibitory concentrations (≥8 μg/mL) showed mutations both in gyrA and parC. The difference in the proportion of strains showing mutations in gyrA and parC was found to be significant (P0.001). The mutation Asp-95→Asn was restricted to Pune strains only. These results indicate that mutations in quinolone target enzymes may have resulted in the high-level resistance seen in these isolates.

BACKGROUND In India, genital ulcer disease (GUD) syndrome is clinically classified as herpetic or nonherpetic and managed accordingly; laboratory support is unavailable at most health facilities. We undertook a study to determine the etiology of GUDs in men presenting to sexually transmitted infection (STI) clinics and assess the performance of the national algorithm for syndromic management of herpetic and nonherpetic GUDs in India. METHOD A cross-sectional study was conducted among men with complaints of genital ulcers attending 8 STI clinics in 4 states. Ulcer swabs were collected and tested by the multiplex polymerase chain reaction method to determine the etiology of GUD. RESULTS Of the 194 men recruited, etiology was confirmed in 121 GUD cases (62%). Herpes simplex virus (48%) was the most common etiological agent identified, followed by Treponema pallidum (23%) and mixed infections (9%). One case of Haemophilus ducreyi was confirmed in this series. The overall sensitivity and specificity of the national syndromic management algorithm for GUD were 68% and 52%, respectively. Using the national algorithm, 52 (42%) cases clinically misclassified as either herpetic (18 cases) or nonherpetic (34 cases) GUD resulting in incorrect treatment. CONCLUSIONS Our findings suggest a revision of existing national STI treatment guidelines in India to include treatment of syphilis infections of all GUD patients. Periodic studies are required to monitor changing spectrum of GUD etiologies in India.

Background Female sex workers (FSWs) in India are provided a standardised package of clinical interventions for management of sexually transmitted infections (STIs). A study was conducted among FSWs at known high STI prevalence sites to determine the effectiveness of the service package. Methods A cohort of FSW clinic attendees in two cities, Hyderabad and Mumbai, were enrolled and followed up from October 2008 to November 2009. At each visit, behavioural and clinical data were obtained and vaginal swabs collected for laboratory testing of cervical infections (gonorrhoea and chlamydia). Results 417 participants were enrolled, of whom 360 attended at least a follow-up visit. Prevalence of cervical infections did not change between the baseline and final visits (27.7% and 21.3% respectively, p=0.08) in spite of presumptive treatment at baseline and syndromic management at all visits. The proportion of asymptomatic cervical infections increased from 36% at baseline to 77% at the final visit. Incidence rate of cervical infections was high (85.6/100 person years) and associated with a prevalent cervical infection at baseline (HR=2.7, p

Biofilm formation is implicated as a potential virulence factor in Candida species and carries important clinical repercussions because of their increased resistance to antifungal treatment, ability to withstand host defences and to serve as a reservoir for continuing infections. The present study was undertaken to determine the biofilm production among oral Candida isolates from HIV-positive and HIV-negative individuals from Pune, India. Biofilm formation was determined using the spectrophotometric or microtitre plate method in 182 Candida isolates, of which 154 were from HIV-positive and 28 were from HIV-negative individuals. A total of 63.2% of the Candida isolates were biofilm producers. Significantly increased biofilm forming abilities both qualitatively as well as quantitatively were observed in Candida isolates from HIV-positive individuals (66.2%) compared to isolates from HIV-negative ones (46.4%), (P- 0.041). Eighty-one (59.6%) C. albicans isolates and 34 (73.9%) non -C. albicans Candida (NCAC) showed biofilm positivity. The NCAC showed significantly greater intensity of biofilm formation compared to the C. albicans, P- 0.032. Our results thus show the enhanced biofilm forming abilities of oral Candida isolates from HIV-infected individuals compared to HIV-uninfected ones and highlight the important role played by biofilm formation in the pathogenesis of NCAC isolates.

The objectives of this cross-sectional study were to determine the prevalence of HIV and sexually transmitted infections (STI) in Hijras (self-identified transgenders of South Asia), study associated risk factors, and compare the prevalence with that in heterosexual men and men having sex with men (MSM) in Pune, India, between 1993 and 2002.After informed consent, individuals attending 3 STI clinics were administered a questionnaire regarding their demographic, socioeconomic, and sexual behaviors. Blood samples were collected for STI and HIV diagnosis. Bivariate and multivariate analyses were performed to determine the correlates of HIV infection.The prevalence of HIV (45.2% in Hijras vs 20% in heterosexual men vs 18.9% in MSM, P0.0001) and warts (10.3% vs 4.6% vs 7.0%; P = 0.004) was higher in Hijras as compared with heterosexual men and MSM; whereas that of genital ulcer disease (15.3% vs 32.6% vs 21.5%; P0.0001) and discharge (5.4% vs 13.6% vs 9.0%; P0.0001) was lower. Hijras were more likely to have received money for sex and have an earlier sexual debut than the comparison groups. In multivariate analysis, receiving money for sex (adjusted odds ratio: 4.49; P0.04) and having genital ulcer disease (odds ratio: 3.87; P0.08) were independently associated with high HIV prevalence in Hijras.Considering the high HIV and STI burden, it is important to review current prevention strategies and stress the need to engage Hijra community members through appropriate targeted intervention programs.

Identification of vulnerability in the HIV-1 envelope (Env) will aid in Env-based vaccine design. We recently found an HIV-1 clade C Env clone (4–2.J45) amplified from a recently infected Indian patient showing exceptional neutralization sensitivity to autologous plasma in contrast to other autologous Envs obtained at the same time point. By constructing chimeric Envs and fine mapping between sensitive and resistant Env clones, we found that substitution of highly conserved isoleucine (I) with methionine (M) (ATA to ATG) at position 424 in the C4 domain conferred enhanced neutralization sensitivity of Env-pseudotyped viruses to autologous and heterologous plasma antibodies. When tested against monoclonal antibodies targeting different sites in gp120 and gp41, Envs expressing M424 showed significant sensitivity to anti-V3 monoclonal antibodies and modestly to sCD4 and b12. Substitution of I424M in unrelated Envs also showed similar neutralization phenotype, indicating that M424 in C4 region induces exposure of neutralizing epitopes particularly in CD4 binding sites and V3 loop.

Recent research has suggested that human papillomavirus (HPV) testing is a superior strategy than the most widely used method, viz., cervical cytology (Pap smears) for age appropriate screening for cervical cancer.1,2 However, HPV testing is still very costly in most developing countries, which bear more than 80% of the global burden of HPV-induced invasive cervical cancer (ICC).3,4 Public health and medical professionals worldwide earnestly await the introduction and wide availability of low-cost, rapid and easily implementable HPV testing methods.5,6 In the interim, however, it is imperative to evaluate and consider implementing cheaper and more practical modalities than cytology for cervical cancer screening, especially for women at high risk of cervical disease. Human immunodeficiency virus (HIV)-infected women constitute one of the highest risk population subgroups for increased incidence and rapid progression of HPV-induced cervical intraepithelial neoplasia (CIN) and ICC.7 India faces large burdens of HIV/acquired immunodeficiency syndrome (AIDS; third highest number of HIV-infected persons in any single nation in the world, with more than a million HIV-infected women) and ICC (almost 30% of the global burden with more than 130,000 new cases and 75,000 deaths annually).4,8 Although HIV-infected women in India and elsewhere continue to live longer due to improved access to antiretroviral therapy, most women lack access to adequate cervical cancer prevention services.9 Visual inspection after application of dilute (3–5%) acetic acid on the cervix (VIA) has been widely researched as an alternative to cytology for use in resource-limited settings.10,11 However, there are very few studies reporting test accuracy of VIA, without verification bias or with biologically salient diagnostic endpoints, among HIV-infected women.12–14 Given this research gap and the need for providing better models for preventive clinical care for HIV-infected women in resource-limited settings, we conducted a cross-sectional study to rigorously evaluate the clinical accuracy of VIA and cytology among HIV-infected women in Pune, India.

Summary Background Opportunistic infections (OIs) influence the morbidity and mortality due to HIV infections. Data from India on the incidence of OIs among HIV-infected individuals by stages of immunodeficiency are scarce. Methods Between September 2002 and November 2004, HIV-infected individuals were enrolled in a prospective study in Pune. They were clinically and immunologically evaluated quarterly. Incidence rates of specific OIs were calculated. Results Median CD4 counts in HIV-infected male and female patients at baseline were 197/mm 3 and 413/mm 3 , respectively. Tuberculosis was the most common OI with an incidence of 15.4 (95% CI 12.2–19.2) per 100 person-years, followed by oral candidiasis 11.3 (95% CI 8.6–14.5), herpes zoster 10.1 (95% CI 7.6–13.1), and cryptococcal meningitis 1.7 (95% CI 0.8–3.1) per 100 person-years. Patients with baseline CD4 counts of 3 were six times more likely to develop OIs compared to those with CD4 counts of >350/mm 3 ( p Conclusions The high incidence of commonly reported OIs in Indian HIV-infected individuals highlights the need for early screening and also the need to increase awareness in healthcare providers, in order to improve decisions regarding prophylaxis for prevention and appropriate therapeutic intervention. Emphasis needs to be given to the early diagnosis and management of tuberculosis in HIV-infected individuals.

A recent report suggesting declining HIV transmission rates in southern India has been based on HIV seroprevalence data to estimate HIV incidence. We analyzed HIV incidence rates among 3 cohorts (male female non-sex worker female sex worker [FSW]) presenting to sexually transmitted infection (STI) clinics in Pune India over 10 years. Between 1993 and 2002 consenting HIV-uninfected individuals were enrolled in a prospective study of the risks for HIV seroconversion. Standardized HIV incidence estimates were calculated separately for the 3 cohorts. HIV acquisition risk declined by more than 70% for FSWs (P = 0.02) and men (P < 0.001) attending the STI clinics. There was no significant reduction in HIV incidence among women attending STI clinics (P = 0.74). The decline in HIV acquisition risk among male patients with STIs was associated with an increase in reported condom use with recent FSW contact and a decrease in genital ulcer disease. We report the first direct evidence for a decline in HIV incidence rates in FSWs and male patients with STIs over time. The lack of change in HIV infection risk among non-sex worker women highlights the need for additional targeted HIV prevention interventions. (authors)

OBJECTIVES: The objectives of this study were to identify sexually transmitted infection (STI) prevalence, assess behavioral and symptom correlates, and develop intervention strategies. GOAL: The goal of this study was to conduct one of the first community-based surveys of STI prevalence and risk behaviors among married men in India. STUDY DESIGN: In 2003, 2,408 randomly selected married men, aged 21 to 40 years, were administered a survey instrument with urine and blood samples collected from a random subset of 641. RESULTS: The most common current STI was gonorrhea (3.9%) with 6.1% of men being positive for an acute STI and 9.7% antibody-positive for Treponema pallidum or herpes simplex virus type 2. Risk behaviors were not associated with laboratory confirmed STIs, but did show an association with men's concerns about sexual performance derived from traditional Indian systems of medicine. CONCLUSION: Culturally based symptoms can serve as effective markers for men involved in risky sexual behaviors and provide an opportunity to engage these men as they seek care for these symptoms at community-based service points.

Resistance to azole antifungals is a significant problem in Candida albicans. An understanding of resistance at molecular level is essential for the development of strategies to tackle resistance and rationale design of newer antifungals and target-based molecular approaches. This study presents the first evaluation of molecular mechanisms associated with fluconazole resistance in clinical C.albicans isolates from India. Target site (ERG11) alterations were determined by DNA sequencing, whereas real-time PCRs were performed to quantify target and efflux pump genes (CDR1, CDR2, MDR1) in 87 [Fluconazole susceptible (n = 30), susceptible-dose dependent (n = 30) and resistant (n = 27)] C.albicans isolates. Cross-resistance to fluconazole, ketoconazole and itraconazole was observed in 74.1% isolates. Six amino acid substitutions were identified, including 4 (E116D, F145L, E226D, I437V) previously reported ones and 2 (P406L, Q474H) new ones. CDR1 over-expression was seen in 77.7% resistant isolates. CDR2 was exclusively expressed with CDR1 and their concomitant over-expression was associated with azole cross-resistance. MDR1 and ERG11 over-expression did not seem to be associated with resistance. Our results show that drug efflux mediated by Adenosine-5'-triphosphate (ATP)-binding cassette transporters, especially CDR1 is the predominant mechanism of fluconazole resistance and azole cross-resistance in C. albicans and indicate the need for research directed towards developing strategies to tackle efflux mediated resistance to salvage azoles.

Background: Oral therapy for pulmonary tuberculosis (TB) treatment suffers from the limitation of hepatic metabolism leading insufficient concentration of antitubercular (anti-TB) drugs in alveolar macrophage which harbors Mycobacterium tuberculosis (MTB). Targeted aerosol delivery of antituberculous drug to lung is efficient for treating local lung TB infection. Objective: The present study was aimed to evaluate rifapentine (RPT) loaded proliposomal dry powder for inhalation (RLDPI) for anti-TBactivity and cytotoxicity in vitro. In vivo toxicity study was also undertaken in Wistar rats to determine safe concentration of RLDPI for administration. Materials and Methods: Anti-TB activity of developed RLDPI was assessed using drug susceptibility testing (DST) on Mycobacteria growth indicator tube (MGIT) method. In vitro cytotoxicity was performed in A549 cell lines and IC 50 values were used to compare the cytotoxicity of formulation with pure RPT. In vivo repeated dose toxicity study was undertaken using Wistar rats at three different doses for 28-days by intratracheal insufflations method. Results: The results of DST study revealed sensitivity of tubercle bacteria to RLDPI at concentration equivalent to 10 μg/mL of RPT. This study confirmed anti-TB potential of RPT in spray-dried RLDPI, though the spray drying method is reported to reduce activity of drugs. Cytotoxicity study in A549 cells demonstrated that RPT when encapsulated in liposomes as RLDPI was safe to cells as compared to pure RPT. In vivo toxicity study revealed that RPT in the form of RLDPI was safe at 1 and 5 mg/kg dose. However, mortality was seen at higher dose (10 mg/kg), possibly because of liver and kidney damage. Conclusion: Thus, these studies demonstrated safety of RLDPI for the treatment of pulmonary TB.

Measurement of C-reactive protein, a marker of inflammation, in dried blood spots has been increasingly incorporated in community-based social surveys internationally. Although the dried blood spot based CRP assay protocol has been validated in the United States, it remains unclear whether laboratories in other less developed countries can generate C-reactive protein results of similar quality. We therefore conducted external quality monitoring for dried blood spot based C-reactive protein measurement for the Indonesia Family Life Survey and the Longitudinal Aging Study in India. Our results show that dried blood spot based C-reactive protein results in these two countries have excellent and consistent correlations with serum-based values and dried blood spot based results from the reference laboratory in the United States. Even though the results from duplicate samples may have fluctuations in absolute values over time, the relative order of C-reactive protein levels remains similar and the estimates are reasonably precise for population-based studies that investigate the association between socioeconomic factors and health.

We assessed the extent of anxiety/depression/distress using Hospital Anxiety and Depression Scale (HADS) among a cross-section of HIV test-seekers at a Voluntary Counseling and Testing (VCT) facility in Pune, India.HADS has 14 items for uniscale with 7 items each for anxiety and depression rated on a four-point Likert scale. Between September 2002 and March 2003, HADS was administered to 150 consecutive HIV tests-seekers attending NARI-Talera VCT facility. Subsequently, HIV testing was done after obtaining informed consent.HADS showed strong internal consistency (Cronbach-alpha 0.77). The prevalence of risk behavior (73.3%) and HIV (45.5%) were high. Education levels influenced anxiety (p = 0.033; 0.008), more so in women (p = 0.044). Repeat test-seekers exhibited significant depression (AOR: 2.9; 95% CI: 1.4-6.1; p = 0.004) and distress (AOR: 2.5; 95% CI: 1.2-5.3; p = 0.017). Marital status influenced the uniscale scores. The HIV positive repeat test-seekers were more anxious (p = 0.035) and depressed (0.037).Existence of emotional distress among HIV test-seekers, particularly among repeat test-seekers, possibly 'AIDS-anxious' individuals indicates additional counseling needs specifically by introducing gender and education sensitive interventions. VCT staff can be trained to assess emotional distress among HIV test-seekers to formulate long-term intervention.

The objective was to investigate changes over a decade in prevalence and correlates of HIV among high-risk women attending sexually transmitted infection (STI) clinics in Pune India who deny a history of commercial sex work (CSW). Design: Cross-sectional. Methods: From 1993 to 2002 2376 women attending 3 STI clinics in Pune were offered HIV screening. Women who denied CSW were included (n = 1020). Of 1020 women 21% were HIV infected. The annual HIV prevalence increased from 14% in 1993 to 29% in 2001-2002 (P < 0.001). The change in HIV prevalence over time was paralleled by changes in clinic visitor characteristics; in later periods women were older more often employed less likely to be currently married and more likely to report condom use. In multivariate analysis factors independently associated with HIV were calendar period (adjusted odds ratio [AOR] 1.9 for 1997-1999 vs. 1993-1996; 95% CI 1.2- 3.0; AOR 2.3 for 2000-2002 vs. 1993-1996; 95% CI 1.5-3.6) lack of formal education (AOR 2.0; 95% CI 1.4-2.9) having been widowed (AOR 3.1; 95% CI 1.6-6.1) current employment (AOR 1.8; 95% CI 1.2-2.6) and genital ulcer disease on examination (AOR 1.8; 95% CI 1.2-2.7). Women attending STI clinics in India who deny a history of CSW represent a small hidden subgroup likely put at risk for HIV because of high-risk behavior of their male partners generally their husbands. Educational and awareness efforts that have targeted other subgroups in India (men and CSWs) should also focus on these hard-to-reach women. Risk reduction in this subgroup of Indian women would also be expected to reduce perinatal infections in India. (authors)

We quantified vaginal lactobacilli and determined their relationship with genital HIV-1 shedding and found a significant negative association between reduced quantity of lactobacilli and cervical HIV-1 viral load (r(2) = - 0.8900, P < 0.01), which may have implications of increased chances of sexual transmission of HIV-1 and genital infections.

To estimate the impact of prevalent and incident herpes simplex virus type 2 (HSV-2) infection on the acquisition of human immunodeficiency virus type 1 (HIV-1), stored serum samples from a cohort of 2732 HIV-1-seronegative patients attending 3 sexually transmitted infection clinics and 1 reproductive tract infection clinic in Pune, India, were screened for HSV-2-specific antibodies. Incident HSV-2 infection was defined serologically as "recent" if a negative result of testing for HSV-2 could be documented within the previous 6 months or "remote" if >6 months had elapsed since the last negative test result. The prevalence of HSV-2 at enrollment was 43%. The HSV-2 incidence was 11.4 cases/100 person-years, and the HIV-1 incidence was 5.8 cases/100 person-years. The adjusted hazard ratios of HIV-1 acquisition from exposure to HSV-2 infection were 1.67 for prevalent HSV-2, 1.92 for remote incident HSV-2, and 3.81 for recent incident HSV-2. Recent incident HSV-2 infection was associated with the highest risk of HIV-1 in this study, which suggests that prevention of HSV-2 infection may reduce the risk of HIV-1 acquisition.

Objective Efforts to prevent HIV transmission from mother to infants in settings like India may benefit from the availability of reliable methods for rapid and simple HIV screening. Data from India on the reliability of rapid HIV test kits are limited and there are no data on the use of rapid HIV tests for screening of pregnant women. Methods Pregnant women attending an antenatal clinic and delivery room in Pune agreed to participate in an evaluation of five rapid HIV tests, including (a) a saliva brush test (Oraquick HIV-1/2, Orasure Technologies Inc.), (b) a rapid plasma test (Oraquick HIV-1/2) and (c) three rapid finger prick tests (Oraquick HIV-1/2; HIV-1/2 Determine, Abbott; NEVA HIV-1/2 Cadila). Results of the rapid tests were compared with three commercial plasma enzyme immunoassay (EIA) tests (Innotest HIV AB EIA, Lab systems/ELISCAN HIV AB EIA, UBI HIV Ab EIA). Results Between September 2000 and October 1, 2001, 1258 pregnant women were screened for HIV using these rapid tests. Forty-four (3.49%) of the specimens were HIV-antibody-positive by at least two plasma EIA tests. All of the rapid HIV tests demonstrated excellent specificity (96-100%). The sensitivity of the rapid tests ranged from 75-94%. The combined sensitivity and specificity of a two-step algorithm for rapid HIV testing was excellent for a number of combinations of the five rapid finger stick tests. Conclusion In this relatively low HIV prevalence population of pregnant women in India, the sensitivity of the rapid HIV tests varied, when compared to a dual EIA algorithm. In general, the specificity of all the rapid tests was excellent, with very few false positive HIV tests. Based upon these data, two different rapid HIV tests for screening pregnant women in India would be highly sensitive, with excellent specificity to reliably prevent inappropriate use of antiretroviral therapy for prevention of vertical HIV transmission.

To determine if the early immunological and virological events of HIV infection are unique in a setting with limited access to health care and HIV-1 subtype C infection, we undertook a prospective cohort study to characterize the early natural history of HIV viral load and CD4(+) T lymphocyte counts in individuals with recent HIV seroconversion in India. CD4(+) T lymphocyte counts were prospectively measured for up to 720 days in 46 antiviral drug-naive persons with very early HIV infection, documented by HIV antibody seroconversion. HIV viral RNA levels were measured subsequently on reposited plasma samples from these same time points. The median viral load "set point" for Indian seroconverters was 28,729 RNA copies/ml. The median CD4(+) cell count following acute primary HIV infection was 644 cells/mm(3). Over the first 2 years since primary infection, the annual rate of increase in HIV viral load was +8274 RNA copies/ml/year and the annual decline in CD4 cell count was -120 cells/year. Although the viral "set point" was similar, the median trajectory of increasing viral load in Indian seroconverters was greater than what has been reported in untreated HIV seroconverters in the United States. These data suggest that the more rapid HIV disease progression described in resource-poor settings may be due to very early virological and host events following primary HIV infection. A rapid increase in viral load within the first 2 years after primary infection may have to be considered when applying treatment guidelines for antiretroviral therapy and opportunistic infection prophylaxis.

Objectives: To estimate the prevalence and incidence of hepatitis B virus (HBV) infection among patients attending three STD clinics in Pune, India, and to identify associated risk factors. Methods: Of the 2098 patients screened at STD clinics in Pune during 1996, 497, who returned for at least one follow up visit, were screened for various markers of HBV infection (HBsAg, anti-HBs, antiHBc), HIV antibody, and VDRL. Results: Of the 497 participants 3.6%, 26.5%, and 43.2% were positive for HBsAg, anti-HBs, and anti-HBc respectively. Tattooing (AOR 1.64, 95% CI 1.03 to 2.64) was found to be independently associated with presence of core antibody. Additionally, history of being in commercial sex work and history of a genital ulcer were independently associated with a positive anti-HBc antibody test (AOR 12.45, 95% CI 5.58 to 27.82 and AOR 1.70, 95% CI 1.09 to 2.66, respectively). 72 out of 497 (14.5%) participants were HIV positive at baseline. HIV-1 antibody positive patients were more likely to have a positive anti-HBc test (69.4% v 39.0%, p

Sir, Intestinal microsporidiosis is the most common cause of chronic diarrhoea in treatment-naive HIV infected patients1. Intestinal microsporidiosis due to Encephalitozoon (Septata) intestinalis and Enterocytozoon bieneusi are also most frequently reported among immunocompromised patients such as transplant recipients2–4. Laboratory diagnosis of microsporidiosis was initially based on the detection of microsporidial spores in stool specimens by light microscopy using special stains5. Electron microscopy, wherever possible, was also used for detection of microsporidia6–8. Diagnosis of intestinal microsporidiosis has been markedly improved by the use of Weber's chromotrope stain and its modified procedure1,5,9, and fluorescence staining with fluorochromes such as Uvitex 2B or calcoflour white which binds to the chitin in the endospore layer of the microsporidial spore1,8,10,11. Molecular technique such as polymerase chain reaction (PCR) became available later for confirmation of micropsoridia. Additionally, species specific PCR can be used to identify different species of microsporidia12–16. However, due to resource constraints, most laboratories rely on the use of microscopy of stool samples for diagnosis of intestinal microsporidiosis. Limited studies have been carried out in India on detection of microsporidia8,17,18. We compared light microscopy with modified trichome stain, fluorescence microscopy using Uvitex 2B and PCR to detect microsporidia in HIV infected individuals with diarrhoea during January 2009 to May 2010 on consecutive HIV infected patients with diarrhoea admitted at Naidu Municipal Corporation Hospital, Pune. The study protocol was approved by the institutional ethics committee & Naidu Municipal Corporation Hospital. The study included 331 patients [65 (51 males, 14 females)] HIV infected adults >18 yr and 266 HIV (uninfected individuals) with diarrhoea. This study was carried out only on HIV infected individuals with diarrhoea. Study patients were interviewed using the structured questionnaire. Diarrhoea was defined as two or more liquid or three or more soft stools per day All the patients had history of diarrhoea of

The present study was undertaken to detect Pneumocystis jirovecii infection among HIV-positive patients presenting with symptoms of lower respiratory tract infection and analyze the associated dihydropteroate synthase (DHPS) and dihydrofolate reductase (DHFR) mutations. P. jirovecii infection was detected in 12.6 % cases. We did not find DHPS gene mutations at the commonest positions of codon 55 and 57; however, mutation at codon 171 was detected in two cases. No mutations in DHFR gene were detected. The results indicate low prevalence of DHPS and DHFR mutations in Indian P. jirovecii isolates, suggesting that the selective pressure of sulfa drugs on the local strains has probably not reached the levels found in developed nations.