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Objective: To evaluate maternal glucose levels during pregnancy as a predictor of adverse perinatal outcomes in Dar es Salaam, Tanzania., Methods: Random blood glucose measurements were analyzed from 3383 pregnant women enrolled in a randomized trial to assess the impact of multivitamins on pregnancy outcomes in Dar es Salaam between August 2001 and July 2004. Information on maternal and neonatal morbidity was recorded at monthly study visits, delivery, and 6 weeks postpartum. Binomial regression and generalized estimating equations were used to determine the relationship between elevated glucose (>7.8 mmol/L) and pregnancy outcomes., Results: In total, 25 women had elevated glucose (0.7%). Hyperglycemia was associated with an increased risk of delivery before 37 weeks [relative risk (RR), 2.11; 95% confidence interval [CI], 1.07-4.13; P=0.03), delivery before 34 weeks (RR, 4.15; 95% CI, 1.43-12.03, P=0.009), incident gestational hypertension (RR, 2.90; 95% CI, 1.24-6.76; P=0.01), low birth weight (RR, 2.87; 95% CI, 1.18-6.99; P=0.02), reduced newborn head circumference (mean difference, -1.57; 95% CI, -2.51 to -0.62; P=0.001), and fetal loss (RR, 3.38; 95% CI, 1.13-10.08; P=0.03)., Conclusion: Maternal hyperglycemia is uncommon among pregnant Tanzanian women, but nonetheless seems to increase the risk of several adverse perinatal outcomes., (Copyright © 2014 International Federation of Gynecology and Obstetrics. Published by Elsevier Ireland Ltd. All rights reserved.)

Background: The effect of dietary macronutrient composition on the rate of gestational weight gain among women in sub-Saharan Africa is unclear., Objective: To examine the effect of macronutrient intake on the rate of gestational weight gain among HIV-negative women in Tanzania., Methods: The weights of 8,428 women were measured monthly from 12 weeks of gestation to term. Prenatal dietary intake was estimated as the cumulative average of multiple 24-hour dietary recalls. The association between energy intake and percentage of energy from carbohydrate, protein, and total fat and rate of weight gain (grams per month) was estimated from generalized estimating equation models. Macronutrient effects were adjusted for total energy using the nutrient density model and maternal age, maternal height, maternal mid-upper-arm circumference, parity, marital status, maternal occupation, maternal education, household wealth, season, and treatment regimen assignment. Body mass index (BMI) was considered as a confounder and a potential modifier of the effect of macronutrient intake on gestational weight gain., Results: A 6 g/month increase in rate of weight gain was associated with every 100-kcal increment in daily total energy intake (95% CI, 1 to 12; p = .03). Analyses substituting 5% of energy from fat by protein showed that weight gain would decrease by 72 g/month (95% CI, 6 to 140; p = .03); substituting 5% of energy from carbohydrate by protein decreased gain by 70 g/month (95% CI, 15 to 124; p = .01). Baseline BMI did not modify these associations., Conclusions: Further research on the effects of macronutrient composition on gestational weight gain is needed to inform the design of supplementation programs for women in developing countries.

Background: Our objectives were to examine the associations of neonatal and infant mortality with preterm birth and intrauterine growth restriction (IUGR), and to estimate the partial population attributable risk per cent (pPAR%) of neonatal and infant mortality due to preterm birth and IUGR., Methods: Participants were HIV-negative pregnant women and their infants enrolled in Dar es Salaam, Tanzania. Gestational age calculated from date of last menstrual period was used to define preterm, and small for gestational age (SGA) was used as proxy for IUGR. Survival of infants was ascertained at monthly follow-up visits. Cox proportional hazard models were used to estimate the associations of preterm and SGA with neonatal and infant mortality., Results: Study included 7225 singletons, of whom 15% were preterm and 21% were SGA; majority of preterm or SGA babies had birthweight ≥2500 g. Compared to term and appropriately sized babies (AGA), relative risks (RR) of neonatal mortality among preterm-AGA was 2.6 [95% CI 1.8, 3.9], RR among term-SGA was 2.3 [95% CI 1.6, 3.3], and the highest risk was among the preterm-SGA babies (RR 15.1 [95% CI 8.2, 27.7]). Risk associated with preterm was elevated throughout the infancy, and risk associated with SGA was elevated during the neonatal period only. The pPAR% of neonatal mortality for preterm was 22% [95% CI 17%, 26%] and for SGA it was 26% [95% CI 16%, 36%]., Conclusions: Preterm and SGA birth substantially increased the risk of mortality. Interventions for prevention and management of these conditions are likely to reduce of infant mortality in Tanzania., (© 2013 John Wiley & Sons Ltd.)

Objectives: Vitamin D is an immunomodulator and can alter response to tuberculosis (TB) treatment, though randomised trials have been inconclusive to date. We present one of the first comprehensive analysis of the associations between vitamin D status and TB treatment, T-cell counts and nutritional outcomes by HIV status., Design: Cohort study., Setting: Outpatient clinics in Tanzania., Participants: 25-hydroxyvitamin D levels were assessed in a cohort of 677 patients with TB (344 HIV infected) initiating anti-TB treatment at enrolment in a multivitamin supplementation (excluding vitamin D) trial (Clinicaltrials.gov identifier: NCT00197704)., Primary and Secondary Outcome Measures: Information on treatment outcomes such as failure and relapse, HIV disease progression, T-cell counts and anthropometry was collected routinely, with a median follow-up of 52 and 30 months for HIV-uninfected and HIV-infected patients, respectively. Cox and binomial regression, and generalised estimating equations were used to assess the association of vitamin D status with these outcomes., Results: Mean 25-hydroxyvitamin D concentrations at enrolment were 69.8 (±21.5) nmol/L (27.9 (±8.6) ng/mL). Vitamin D insufficiency (<75 nmol/L) was associated with a 66% higher risk of relapse (95% CI 4% to 164%; 133% higher risk in HIV-uninfected patients). Each unit higher 25-hydroxyvitamin D levels at baseline were associated with a decrease of 3 (p=0.004) CD8 and 3 (p=0.01) CD3 T-cells/µL during follow-up in patients with HIV infection. Vitamin D insufficiency was also associated with a greater decrease of body mass index (BMI; -0.21 kg/m(2); 95% CI -0.39 to -0.02), during the first 8 months of follow-up. No association was observed for vitamin D status with mortality or HIV disease progression., Conclusions: Adequate vitamin D status is associated with a lower risk of relapse and with improved nutritional indicators such as BMI in patients with TB, with or without HIV infection. Further research is needed to determine the optimal dose of vitamin D and effectiveness of daily vitamin D supplementation among patients with TB.

Many studies have documented a high prevalence of anemia among tuberculosis (TB) patients and anemia at TB diagnosis has been associated with an increased risk of death. However, little is known about the factors contributing to the development of TB-associated anemia and their importance in TB disease progression. Data from a randomized clinical trial of micronutrient supplementation in patients with pulmonary TB in Tanzania were analyzed. Repeated measures of anemia with iron deficiency, anemia without iron deficiency, and iron deficiency without anemia were assessed as risk factors for treatment failure, TB recurrence, and mortality. The prevalence of anemia (hemoglobin < 110 g/L) at baseline was 64%, more than one-half of which was related to iron deficiency (mean corpuscular volume , 80 fL). We found no evidence of an association between anemia (with or without iron deficiency) or iron deficiency without anemia at baseline and the risk of treatment failure at 1 mo after initiation. Anemia without iron deficiency was associated with an independent, 4-fold increased risk of TB recurrence [adjusted RR = 4.10 (95% CI = 1.88, 8.91); P < 0.001]. Iron deficiency and anemia (with and without iron deficiency) were associated with a 2- to nearly 3-fold independent increase in the risk of death [adjusted RR for iron deficiency without anemia = 2.89 (95% CI = 1.53, 5.47); P = 0.001; anemia without iron deficiency = 2.72 (95% CI = 1.50, 4.93); P = 0.001; iron deficiency anemia = 2.13 (95% CI = 1.10, 4.11); P = 0.02]. Efforts to identify and address the conditions contributing to TB-associated anemia, including iron deficiency, could play an important role in reducing morbidity and mortality in areas heavily affected by TB.

Background: Human Immunodeficiency Virus type 1 (HIV-1) infection leads to a progressive decline in CD4+ T-lymphocyte (CD4) cells. Initiation of prophylaxis against Opportunistic infections in adults (CD4% used for children) and antiretroviral therapy is usually based on CD4 cell counts, but CD4 cell counts measurement is not affordable in most African countries., Objective: To examine whether total lymphocyte counts (TLC) may be used as proxies for low CD4 cell counts., Design: Cross-sectional at baseline when women were pregnant and at least six months postpartum., Methods: 1,078 HIV-1-infected pregnant women from Dar es Salaam, Tanzania were enrolled in a randomized clinical trial. A series of receiver operator characteristic (ROC) curves were created at baseline and at least 6 months postpartum and among women in WHO Stage 3 and above. The sensitivity and specificity of TLC and hemoglobin in predicting an absolute CD4 count < 200 cells/mm3 were determined for various clinically relevant cut points., Results: TLC was not a good predictor of low CD4 cell counts during pregnancy or at least six months postpartum as exhibited by low ROC Area Under the Curve (AUCs) of .57 and .62 respectively. No other variable had the ability to predict CD4 < 200 cells/mm3., Conclusions: The use of TLC as a proxy for the estimation of low CD4 cell counts in a population of HIV-1-infected adults from sub-Saharan Africa was not substantiated. Inexpensive methods to quantify CD4 cell counts in Africa are needed.

Background: HIV has fuelled the TB epidemic in sub-Saharan Africa. Mortality in patients co-infected with TB and HIV is high. Managing factors influencing mortality in TB patients might help reducing it. This study investigates factors associated with mortality including patients' HIV sero-status, CD4 cell count, laboratory, nutritional and demographic characteristics in AFB smear positive pulmonary TB patients., Methods: We studied 887 sputum smear positive PTB patients, between 18 and 65 years of age receiving standard 8 months anti-TB treatment. Demographic, anthropometric and laboratory data including HIV, CD4 and other tests were collected at baseline and at regular intervals. Patients were followed for a median period of 2.5 years., Results: Of the 887 participants, 155 (17.5%) died, of whom 90.3% (140/155) were HIV-infected, a fatality of 29.7% (140/471) compared to 3.6% (15/416) among HIV-uninfected. HIV infection, age, low Karnofsky score, CD4 cell counts and hemoglobin, high viral load, and oral thrush were significantly associated with high mortality in all patients., Conclusion: Mortality among HIV-infected TB patients is high despite the use of effective anti-TB therapy. Most deaths occur after successful completion of therapy, an indication that patients die from causes other than TB. HIV infection is the strongest independent predictor of mortality in this cohort.

Background: Reference values for hematological and biochemical assays in pregnant women and in newborn infants are based primarily on Caucasian populations. Normative data are limited for populations in sub-Saharan Africa, especially comparing women with and without HIV infection, and comparing infants with and without HIV infection or HIV exposure., Methods: We determined HIV status and selected hematological and biochemical measurements in women at 20-24 weeks and at 36 weeks gestation, and in infants at birth and 4-6 weeks of age. All were recruited within a randomized clinical trial of antibiotics to prevent chorioamnionitis-associated mother-to-child transmission of HIV (HPTN024). We report nearly complete laboratory data on 2,292 HIV-infected and 367 HIV-uninfected pregnant African women who were representative of the public clinics from which the women were recruited. Nearly all the HIV-infected mothers received nevirapine prophylaxis at the time of labor, as did their infants after birth (always within 72 hours of birth, but typically within just a few hours at the four study sites in Malawi (2 sites), Tanzania, and Zambia., Results: HIV-infected pregnant women had lower red blood cell counts, hemoglobin, hematocrit, and white blood cell counts than HIV-uninfected women. Platelet and monocyte counts were higher among HIV-infected women at both time points. At the 4-6-week visit, HIV-infected infants had lower hemoglobin, hematocrit and white blood cell counts than uninfected infants. Platelet counts were lower in HIV-infected infants than HIV-uninfected infants, both at birth and at 4-6 weeks of age. At 4-6 weeks, HIV-infected infants had higher alanine aminotransferase measures than uninfected infants., Conclusion: Normative data in pregnant African women and their newborn infants are needed to guide the large-scale HIV care and treatment programs being scaled up throughout the continent. These laboratory measures will help interpret clinical data and assist in patient monitoring in a sub-Saharan Africa context.

Objective: To determine the demographic, socio-economic and psycho-social factors associated with condom use amongst antenatal clinic attendees in Dar es Salaam., Methods: A cross sectional study design was employed in four antenatal clinics in Dar es Salaam. Pregnant women were interviewed between April 1995 and July 1997 to find out if they have ever used a condom and if so whether they had used them consistently for all coital acts in the previous year., Results: Of 1,585 women interviewed, 41% had their first sexual experience before age of 18 years and 82% had a history of having more than two sexual partners during their lifetime. Sixty-two percent of women had never used a condom. Although 40% had used a condom in the previous year only 12% used them consistently. Ever use of a condom increased significantly with the number of years of education of the respondent and her partner also with the respondent's financial independence. Women with > 9 years of education were twice as likely as women with < 5 years of education to be condom users (prevalence ratio (PR) = 2.1, 95% confidence interval (CI) = 1.6-2.7). Professional women were almost twice as likely as housewives to have ever used a condom (PR = 1.8, 95% CI = 1.3-2.3). Women who reported that they have had more than four sexual partners during their lifetime were associated with nearly a four-fold higher lifetime rate of having ever used a condom, compared with a single lifetime partnership (PR = 3.9, 95% CI = 2.8-5.4)., Conclusion: The reported prevalence of ever use of a condom amongst antenatal clinic attendees is low and inconsistent especially among HIV positive women. Deliberate effort should be used to ensure condom access, availability and correct and consistent use of condoms by women in all sexual acts.

Background: The United Republic of Tanzania is one of the 22 high M. tuberculosis burden countries. Data collected between 2002 and 2007 indicate that the global prevalence of drug-resistant M. tuberculosis including MDR vary greatly. The varied drug-resistance patterns make continuous surveillance of drug resistance an essential component of tuberculosis control program., Findings: M. tuberculosis isolates were obtained from consenting adult tuberculosis patients involved in a placebo-controlled study to evaluate the efficacy of multivitamin supplements on response to anti-Tb treatment in Dar es Salaam, Tanzania. Antimicrobial susceptibility testing was done on four antimicrobial agents namely streptomycin, isoniazid, ethambutol and rifampicin. HIV testing and CD4+ T lymphocytes enumeration were also done. A total of 280 M. tuberculosis isolates from 191 (68%) males and 89 (32%) female patients with no previous history of anti-tuberculosis treatment exceeding 4 weeks in the previous 12 months were tested. Among these, 133 (47%) patients were HIV seropositive. Fourteen (5.0%) isolates were resistant to any of the anti-tuberculosis drugs. The prevalence of primary resistance was 5.0%, 0.7%, 0.4% and 0% for isoniazid, streptomycin, rifampicin and ethambutol respectively. One isolate (0.4%) was MDR, with resistance to isoniazid, streptomycin and rifampicin., Conclusion: M. Tb primary resistance rate in a selected population in Dar es Salaam Tanzania is low and efforts should be undertaken to support the Tuberculosis program.

Background: Tuberculosis (TB) often coincides with nutritional deficiencies. The effects of micronutrient supplementation on TB treatment outcomes, clinical complications, and mortality are uncertain., Methods: We conducted a randomized, double-blind, placebo-controlled trial of micronutrients (vitamins A, B complex, C, and E, as well as selenium) in Dar es Salaam, Tanzania. We enrolled 471 human immunodeficiency virus (HIV)-infected and 416 HIV-negative adults with pulmonary TB at the time of initiating chemotherapy and monitored them for a median of 43 months., Results: Micronutrients decreased the risk ofTB recurrence by 45% overall (95% confidence interval [CI], 7% to 67%; P = .02) and by 63% in HIV-infected patients (95% CI, 8% to 85%; P = .02). There were no significant effects on mortality overall; however, we noted a marginally significant 64% reduction of deaths in HIV-negative subjects (95% CI, -14% to 88%; P = .08). Supplementation increased CD3+ and CD4+ cell counts and decreased the incidence of extrapulmonary TB and genital ulcers in HIV-negative patients. Micronutrients reduced the incidence of peripheral neuropathy by 57% (95% CI, 41% to 69%; P < .001), irrespective of HIV status. There were no significant effects on weight gain, body composition, anemia, or HIV load., Conclusions: Micronutrient supplementation could improve the outcome in patients undergoing TB chemotherapy in Tanzania.

Objective: To evaluate C-reactive protein (CRP) as a predictor of HIV-related outcomes among women and children in a resource-poor setting., Design: We measured serum CRP concentration among 606 HIV-infected women, all of whom were not taking highly-active antiretroviral therapy, 3 to 11 months after they gave birth, and assessed relationships of CRP to HIV-related endpoints, including maternal disease progression, mother-to-child transmission of HIV, and maternal and child mortality., Methods: We used Cox proportional hazards and regression models adjusted for age, sociodemographic characteristics, anthropometric measurements, hemoglobin, CD4 cell count, HIV viral load, and, for child outcomes, breastfeeding status., Results: Ninety-four women had a high CRP concentration (> 10 mg/l). During the follow-up, 56 women progressed to WHO stage 4 and 188 died, and a high maternal CRP concentration was associated with a 2.26-fold [95% confidence interval (CI), 1.64-3.12] greater risk of progression to stage 4 or death. Among children, 174 acquired HIV and 116 died by age 2 years, and a high maternal CRP concentration was associated with a 3.03-fold (95% CI, 1.85-4.96) greater risk of child mortality. In multivariate analyses among adults, a high maternal CRP concentration was associated with a 1.55-fold (95% CI, 1.08-2.23) greater risk of progression to stage 4 or death. A maternal CRP concentration was not significantly associated with mother-to-child transmission of HIV., Conclusions: A high maternal CRP concentration independently predicts HIV disease progression, maternal mortality, and child mortality in a resource-poor setting. C-reactive protein may be an important and inexpensive prognostic indicator for HIV-infected women and their children.

The aim of this study was to evaluate the performance of two antibody enzyme-linked immunosorbent assays (ELISAs) [Vironostika Uni-Form II plus O and Enzygnost anti-HIV-1/2 Plus], and two antigen/antibody combination ELISAs [Murex and Vironostika HIV Uni-Form II] for use in an alternative confirmatory HIV diagnostic testing strategy in Dar es Salaam, Tanzania. Altogether, 1380 serum samples were included. All ELISA reactive samples were tested using the Inno-Lia antibody assay and discrepant samples were tested on the Innotest p24 antigen assay. Three hundred and one (21.8%) samples were confirmed HIV-1 antibody positive by Inno-Lia including 27/508 (5.3%) from blood donors, 65/511 (12.7%) from pregnant women and 209/361 (57.9%) from hospital patients. The sensitivity at initial testing was 100% (95% CI; 98.8-100%) for all assays except Vironostika Uni-Form II plus O (99.7%; 95% CI; 98.2-99.9%) which showed one false negative sample at initial testing but 100% sensitivity after repeat testing. The final specificity at repeat testing was 100% (95% CI; 99.7-100%) for Enzygnost anti-HIV-1/2 Plus, 99.4% (95% CI; 98.8-99.8%) for each of the antigen/antibody combination ELISAs and 97.9% (95% CI; 96.8-98.6%) for Vironostika plus O ELISA. An alternative confirmatory HIV testing strategy based on initial testing on any of the two antigen/antibody assays followed by testing of reactive samples on the Enzygnost anti-HIV-1/2 Plus assay gave 100% specificity (95% CI; 99.7-100%).

Background: Sputum microscopy for acid-fast bacilli (AFB) is the commonest diagnostic method for pulmonary tuberculosis (PTB) in developing countries. The method is reported to be less sensitive in human immunodeficiency virus (HIV) positive compared to negative patients. We determined the bacillary density in sputum of smear-positive PTB patients and related it to the patients' HIV status, CD4 cell count, clinical and demographic characteristics., Methods: Three sputum samples per patient were examined using microscopy before initiating therapy. The AFB density was graded according to World Health Organization recommendations. The smear with the highest density was used. High bacillary density was defined as >10 AFB/field. HIV status and CD4 cell count were determined according to the national guidelines., Results: Of 844 patients, 433 (51.3%) were HIV-positive. High bacillary density was significantly less common among HIV-positive (39.0%) than -negative (75.7%) patients (prevalence ratio 0.52; 95%CI 0.45-0.59, P < 0.0001). Among HIV-positive patients, the proportion of those with high bacillary density increased progressively with CD4 cell counts (P = 0.003)., Conclusion: HIV is associated with lower AFB concentration in sputum. The AFB density falls with falling CD4 cell count. Microscopy for AFB in sputum may be less sensitive in diagnosing PTB when HIV infection is present, especially in severely immunocompromised patients.

Background: Many developing countries are currently burdened by both undernutrition and increasing rates of overweight and obesity. Scarce data are available from population studies on the recent trends and current epidemiology of obesity in African settings., Objectives: The objectives were to evaluate changes in the prevalence of obesity, underweight, and wasting in women of reproductive age from Dar es Salaam, Tanzania, during the past 10 y and to identify contemporary sociodemographic correlates of these indicators., Design: We estimated the prevalence of obesity [body mass index (BMI; in kg/m2) > or = 30], underweight (BMI < 18.5), and wasting (midupper arm circumference < 22 cm) in 73 689 women aged 14-52 y who attended antenatal care clinics in the city of Dar es Salaam, Tanzania, between 1995 and 2004., Results: The prevalence of obesity rose steadily and progressively from 3.6% in 1995 to 9.1% in 2004 [adjusted prevalence ratio (PR): 1.97; 95% CI: 1.66, 2.33; P for trend for year < 0.0001]. Underweight showed only a modest decline from 3.3% in 1995 to 2.6% in 2004 (adjusted PR: 0.91; 95% CI: 0.75, 1.10; P for trend for year = 0.003), whereas no change was observed in the prevalence of wasting. In the most recent years (2003 and 2004), obesity was positively associated with age, parity, and socioeconomic status and inversely with HIV infection. Underweight was inversely related to socioeconomic status and positively to HIV status., Conclusion: The recent, rapid, and large increase in the prevalence of obesity in women represents a new competing public health priority in urban Tanzania, where underweight and wasting have not decreased substantially.

Background: Syndromic management of STIs has been advocated as simplified and cheap approach. Youth have been reported to be at increased risk of acquiring STIs which can facilitate HIV transmission. We have investigated the relationship between the syndromic management and specific aetiology diagnosis and its relationship with HIV infection and health seeking behaviour among youth attending a reproductive health clinic in Dar es Salaam, Tanzania., Methods: Between September 1998 and February 1999 among 1895 adolescents and youth below 25 years seen in the clinic 199 (10.5%) were randomly selected and consented to participate in the study. A standard questionnaire was administered. Blood and vaginal or urethral specimens were taken and investigated for STI causative agents., Results: Among a total of 199 studied adolescents and youth 22.6 % were teenagers, with fewer females 17.8% than males; 27.5% (p < 0.018). 20.8% of the females compared to 11.5% in males were HIV infected. Genital discharge was the most common complaint which was reported in 54.1% of male and 63.4 % of female patients. All males with gonorrhoea and four out of five with Chlamydia were given appropriate treatment with syndromic management, while 28% women with gonorrhoea or Chlamydia received appropriate treatment by syndromic management. All patients found with active syphilis by serology had not complained of genital ulcers and would not have been assigned to syndromic treatment for syphilis at the initial visit., Conclusion: The burden of STIs in this youth population is large indicating that youth are at increased risk of STIs and will certainly require youth friendly clinics. There is a need to refine the current syndromic management guidelines.

This study aimed at assessing the extent to which decline in HIV infection prevalence reflects decline in incidence in 3 areas with contrasting initial exposure to the HIV epidemic in the Kagera region of Tanzania. A population sample was recruited for the baseline study in 1987 through a multistage cluster sampling technique to determine HIV prevalence. Seronegative individuals identified in the baseline and subsequent studies were followed up for 3 years to yield trends in incidence that were compared for the 3 areas. The overall age-adjusted HIV-1 prevalence in the high-prevalence area of Bukoba urban declined significantly from 24.2% in 1987 to 18.2% in 1993 and later to 13.3% in 1996 (P = 0.0001). In the medium-prevalence area of Muleba, overall age-adjusted prevalence declined significantly from 10.0% in 1987 to 6.8% in 1996 and later to 4.3% in 1999 (P = 0.0003), whereas in the low-prevalence area of Karagwe the prevalence declined from 4.5% in 1987 to 2.6% in 1999 (P = 0.01). In all 3 areas, the most significant decline was consistently observed among women in the age group 15-24 years. No age group exhibited a significant upward prevalence trend. The HIV-1 incidence for Bukoba urban declined from 47.5 to 9.1 per 1000 person-years of observation in 1989 and 1996, respectively, whereas in Muleba it decreased from 8.2 to 3.9 in 1989 and 2000, respectively. Sex-specific estimates indicated a significant decline among women in the high-prevalence area of Bukoba urban from 51.5 to 9.2 per 1000 person-years at risk (P = 0.001). It is concluded that the HIV-1 epidemic in Kagera is on the decrease as reflected by the decline in HIV-1 incidence and prevalence trends particularly among the 15-24 year olds. The decline in the 3 areas of differing magnitude implies that the HIV/AIDS epidemic may be arrested early without necessarily peaking to saturation levels.

Background: Resistance to third generation cephalosporins due to acquisition and expression of extended spectrum beta-lactamase (ESBL) enzymes among Gram-negative bacteria is on the increase. Presence of ESBL producing organisms has been reported to significantly affect the course and outcome of an infection. Therefore infections due to ESBL isolates continue to pose a challenge to infection management worldwide. The aim of this study was to determine the existence and to describe phenotypic and genotypic characteristics of ESBLs in an Intensive Care Unit (ICU) setting in Tanzania., Methods: Between October 2002 and April 2003, clinical information and samples were collected from patients suspected to have nosocomial infections in an Intensive Care Unit of a tertiary hospital in Tanzania. The isolates were identified, tested for antimicrobial susceptibility and analysed for presence of ESBL genes., Results: Thirty-nine Gram-negative bacteria were isolated from clinical samples of 39 patients. These isolates included 13 Escherichia coli, 12 Enterobacter spp, 5 Pseudomonas spp, 4 Proteus spp, 2 Klebsiella. pneumoniae, 2 Citrobacter freundii and 1 Chryseomonas luteola. Eleven (28.2%) of these isolates were ESBL producing. The ESBL genes characterised were SHV-12, SHV-28 and CTX-M-15. The ESBL producing isolates were more resistant to gentamicin and ciprofloxacin than non-ESBL producing isolates., Conclusion: This study shows the presence of ESBL genes among Gram-negative bacteria in the ICU setting in Tanzania. There is a need to institute strict hospital infection control policy and a regular surveillance of resistance to antimicrobial agents.

Malaria infection during pregnancy increases the risk of adverse birth outcomes among HIV-infected women. The role of umbilical cord parasitemia is not well characterized. We examined the risk of adverse perinatal outcomes in relation to maternal or umbilical cord Plasmodium falciparum parasitemia among 275 HIV-infected women from Tanzania, who participated in a randomized trial of zinc supplementation during pregnancy. Maternal parasitemia (> or = 1/microL) at the first antenatal visit was associated with increased risk of low birth weight < 2,500 g (adjusted relative risk [ARR] = 2.66; P = 0.01) and preterm delivery < 37 weeks (ARR = 1.87; P = 0.06). Maternal parasitemia at delivery was associated with preterm delivery (ARR = 2.27; P = 0.008), intrauterine growth retardation (ARR = 1.92; P = 0.03), and neonatal death (ARR = 3.22; P = 0.07). Cord parasitemia was associated with a large and significant increase in the risk of neonatal death (ARR = 8.75; P = 0.003). Maternal parasitemia at the first antenatal visit was strongly related to parasitemia at delivery, and the latter was associated with cord blood parasitemia. CD4 cell counts, parity, or assignment to the zinc arm (25 mg daily) were not associated with parasitemia in maternal or cord blood at delivery. Successful treatment of HIV-infected women who present to the first prenatal visit with malaria parasitemia and avoidance of reinfection are likely to decrease the risk of adverse outcomes during pregnancy and the early postpartum period. Cord blood parasitemia is a strong predictor of neonatal death. The potential effect of zinc supplementation on clinical malaria outcomes deserves future investigation.

Background: In observational studies, the zinc status of HIV-infected persons has been associated with both positive and adverse clinical outcomes. Such endpoints may affect the risk of adverse birth outcomes among HIV-infected women., Objective: We examined the effects of zinc supplements on birth outcomes, hematologic indicators, and counts of T lymphocyte subsets among 400 HIV-infected pregnant women., Design: Eligible women between 12 and 27 wk of gestation were randomly assigned to daily oral supplementation with either 25 mg Zn or placebo between recruitment and 6 wk after delivery. All women received iron, folic acid, and multivitamin supplements irrespective of the experimental assignment., Results: We observed no significant differences in birth weight, duration of gestation, or fetal and neonatal mortality between women in the zinc and placebo groups. Hemoglobin concentrations increased between baseline and 6 wk postpartum in both groups. However, the rise in hemoglobin over this period was significantly lower (P = 0.03) in the zinc group (x +/- SD: 11.5 +/- 17.9 g/L) than in the placebo group (15.2 +/- 18.6 g/L). Similarly, the changes in red blood cell count and in packed cell volume over the same period were significantly lower in the zinc group (P < 0.01 and P = 0.01, respectively). Zinc had no effect on CD4(+), CD8(+), or CD3(+) cell counts during the follow-up period., Conclusion: Because of the lack of beneficial effects of zinc on adverse pregnancy outcomes and the likelihood of negative effects on hemoglobin concentrations, no compelling evidence exists to support the addition of zinc to prenatal supplements intended for pregnant HIV-infected women.

Background: Increased prevention of maternal-to-child transmission of HIV-1 has now become possible due to the availability of effective antiretroviral drugs in developing countries. It is necessary for pregnant women to know their HIV status in order to administer timely treatment to reduce transmission of the virus. This study assesses correlates of acceptance of testing for HIV infection in the antenatal setting in Dar-es-Salaam, Tanzania., Methods: Between August 13, 2001 and November 27, 2002, 14,235 pregnant women were offered screening for HIV as part of routine prenatal care. Demographic information pertaining to the women and their partners, if applicable, was collected. Univariate and multiple logistic regression analyses were carried out., Results: The majority of women were married monogamously (60.0%), had < or =7 years of education (75%), and were unemployed (70.4%). Of the 14,235 women offered screening for HIV, 10,991 (77.2%) accepted. Site of recruitment was significantly associated with screening acceptance (P for trend < 0.0001). Additionally, age, education, marital status, and partner's occupation were significant predictors of testing acceptance., Conclusion: The site at which recruitment occurs is a significant factor in determining a woman's odds of accepting HIV testing. The site covariate includes such factors as individual counselor effects, length of waiting time, and length of time the site has been operational.

Objective: To determine the rate of decline of CD4 T lymphocytes among HIV-1-infected individuals., Design and Setting: A prospective open cohort study of workers in three hotels in Dar es Salaam., Methods: The workers were seen yearly during the study. CD4 T lymphocyte counts were determined using flow cytometry. The CD4 T-lymphocyte slopes were determined using a linear regression model., Results: During the 9-year study period 682 subjects were selected for lymphocyte subset determinations. Of these, 94 HIV-1-seroprevalent (72%), 77 HIV-1-seroincident (67%) and 325 seronegative (75%) individuals had three or more CD4 T-cell determinations, and were used for calculations of CD4 cell slopes with a mean follow-up period of 71.4, 52.9 and 86.0 months, respectively. The median yearly decline of the CD4 T-lymphocyte counts and percentages among seroprevalent individuals was -21.5 cells/microl and -1.3%; among the seroincident individuals the median decline was -22.0 cells/microl and -1.5%. In seroincident individuals the mean duration to a CD4 T-lymphocyte level corresponding to a definition of AIDS was 13.3 years or 11.8 years for CD4 cell counts or percentages, respectively. HIV-1-seropositive subjects who died had significantly steeper CD4 cell slopes than those who survived., Conclusion: The rates of CD4 T-lymphocyte decline in HIV-1-infected individuals in our population are similar to those reported in Europe and north America.

Current HIV management guidelines are based on natural history studies from the developed world. Data on the similarity of the natural course of HIV-1 infection conflict with studies in the developing world. A cohort of 1887 hotel workers with no access to antiretroviral therapy was followed between 1990 and 1998 in Dar es Salaam through annual clinical evaluations and CD4+ T-lymphocyte (CD4 cell) count determinations. 196 (10.4%) were HIV-1 sero-prevalents; 133 (7.9%) were HIV-1 sero-incidents; and 1558 (82.6%) remained HIV seronegative. Follow-up duration was 13,719 and 82,742 months for HIV-1 seropositives and HIV seronegatives respectively. Clinical events occurred at median CD4 cell counts similar to those previously reported from the developed world, but death occurred at higher counts. Off-duty last 6 months, chronic diarrhoea and a faster CD4 cell count decline were associated with faster disease progression and death. In Tanzania HIV natural history is similar to that from the developed world and similar management guidelines could be employed.

The level of CD4(+) T-lymphocytes represents a useful marker with which to monitor the progression of HIV infection. Sex and geographical differences in the reference values of lymphocyte subsets have been reported. We have compared two flow cytometric methods (MultiSET and SimulSET) for the quantification of lymphocyte subsets using whole blood from 92 HIV seropositive and 241 seronegative adults, and determined the reference values of lymphocyte subsets in HIV seronegative Tanzanian subjects. In seronegative Tanzanian subjects, the percentages of CD3(+) and CD4(+) T-lymphocytes and the CD4(+):CD8(+) T-lymphocyte ratios were lower while the percentage of natural killer cells was higher compared to the levels of the corresponding parameters reported for Europeans. Seronegative Tanzanian females had significantly higher levels of CD3(+) and CD4(+) T-lymphocytes and CD4(+):CD8(+) T-lymphocyte ratios compared to seronegative males. The correlation coefficients of CD3(+), CD4(+) and CD8(+) T lymphocyte counts and percentages obtained by the two flow cytometric methods were high. The median values of the number of CD4(+) T-lymphocytes obtained by the two methods were not significantly different. In conclusion, determination of the reference values of lymphocyte subsets in HIV seronegative Tanzanian adults showed significant sex differences and differences in percentage values compared to those reported in certain other geographical areas. There was acceptable agreement in the levels of CD4(+) T-lymphocyte values obtained by the two flow cytometric methods.

Human herpesvirus 8 or Kaposi's sarcoma-associated herpesvirus (HHV8/KSHV) is believed to be the most important etiopathological factor of Kaposi's sarcoma (KS) and some specific types of malignant lymphomas. The diagnostic and prognostic significance of serum viral load in endemic (African) areas is poorly understood. In AIDS-related KS (AKS) it has been shown that HIV-Tat may be of pathogenic importance and that immunoreactivity to Tat may have prognostic significance. Here we report on the quantitative analysis of HHV8 DNA in serum from Tanzanian patients with KS (n = 19), either AIDS-related (AKS) (n = 14) or endemic KS (EKS) (n = 5) and non-KS control individuals (n = 4). Fourteen AKS sera were also tested for HIV-tat antibodies by a direct ELISA assay. In AKS patients detectable (12 out of 14) serum HHV8 DNA levels showed a median of 1400 copies/ml as compared to a median of 200 copies/ml for EKS, but for one AKS case with an exceptionally high level (25,500 copies/ml). The serum HHV8 DNA levels were usually higher in males (n = 17; median 580 DNA copies/ml) as compared to females (n = 6; median 120 DNA copies/ml) and in early, patch stages (n = 8; median 2,750 copies/ml) as compared to late, nodular stages (n = 11; median 200 DNA copies/ml). Of fourteen sera from AKS patients, seven were positive for antibody against HIV-1 tat. Epitope analysis of the anti-tat antibody spectrum showed reactivity to various non-functional sites, but not towards the functional epitopes 46-60 (TAR-binding region).

AIDS-associated Kaposi's sarcoma (AKS) is particularly aggressive and it is one of the principal neoplasms in regions of Africa affected by both high endemic HHV8 and epidemic HIV infection. In this study, serum samples from 18 patients with Kaposi's sarcoma from Tanzania, mostly males (n = 15 vs 3), were subjected to analysis with respect to HHV8-DNA load and antibody spectrum against the HIV-1 tat protein. Of the 18 patients, 14 were HIV-1-positive. The median HHV8 virus load in the HIV-1-positive group was 2075 DNA copies/ml, compared to 450 copies/ml in the HIV-1-negative group. In the HIV-1-positive group, the males had a higher HHV8-DNA virus load as compared to females (median: 4600 vs 1400 genome copies per ml). Since tat can promote AKS development (4-6) by intercellular signalling pathways, and these signals can be abolished by anti-tat IgG (7-9), we have examined the anti-tat IgG spectrum in this study. It would be expected that the levels of serum HHV8-DNA are higher in KS patients who have low anti-tat IgG titer, or who are anti-tat IgG-negative. In the present study, seven out of fifteen AKS patients were positive for anti-tat IgG. Although, we have not seen a strict quantitative relationship between serum anti-tat IgG and HHV8-DNA levels, our data appear to suggest a correlation between the two parameters. In view of these observations and the published data, we suggest that cross-signalling pathways between the tat protein and HHV8-DNA are involved in the complexity of pathogenesis of Kaposi's sarcoma.

Human herpesvirus 8 (HHV-8) is associated with Kaposi's sarcoma. There is a high seroprevalence of HHV-8 in several African countries, but the transmission route is not known definitively. In this study 174 serum samples from blood donors in Tanzania were examined by immunofluorescence assays detecting antibodies to latent and lytic HHV-8 antigens. Real-time polymerase chain reaction was used for detection and quantification of HHV-8 DNA in serum. In all, 83/174 (48%) of the subjects had antibodies to latent or lytic antigens. Forty (23%) had antibodies to both antigens and of those eight (20%) had detectable HHV-8 DNA in serum. HHV-8 DNA load correlated with antibody titres to lytic, but not latent, HHV-8 antigens. This supports the usefulness of anti-lytic antibodies in HHV-8 serology and suggests that transmission of HHV-8 by blood contact could be of importance in this region., (Copyright 2002 Wiley-Liss, Inc.)

This study was conducted to determine the risk factors for recent (active) syphilis among HIV-1 seropositive pregnant women (N = 1058) in Dar es Salaam, Tanzania, Recruitment of study participants (N = 1058) was done between April 1995 and June 1997 at four main prenatal clinics in Dar es Salaam city. Study subjects were interviewed to obtain information about potential risk factors, and blood and genital specimens were collected for detection of syphilis and other genital infections. The prevalence of active syphilis was 5.9%. After adjusting for other risk factors, women without their own source of income had a 50% lower risk of syphilis (OR = 0.5, 95% CI: 0.3-0.9). The risk of active syphilis was significantly increased among women with genital ulceration on examination (OR = 8.4, 95% CI: 1.5-47.7), and in those with trichomoniasis (OR = 2.2, 95% CI: 1.2-3.8). HIV-related immunodeficiency was not associated with increased risk of syphilis. These results show that syphilis and other genital infections are a major problem among HIV infected women. Prevention of syphilis and other genital infections is urgently needed in this population.

Objective: To determine and compare the antimicrobial susceptibility patterns of Vibrio cholerae 01 strains, which were isolated in two cholera epidemics in 1997 and 1999 in Dar es Salaam., Methods: V. cholerae 01 strains isolated from patients with cholera in Dar es Salaam city during 1997 (94 isolates) and 1999 (87 isolates) were stored on nutrient agar slants at room temperature and antimicrobial susceptibility pattern was determined, using Kirby Bauer method., Setting: Department of Microbiology and Immunology, Muhimbili Medical Centre, Dar es Salaam, Tanzania., Results: A total of 181 V. cholerae 01 strains were studied during two epidemic periods when tetracycline or erythromycin was used for treatment of patients with severe disease. Among the 94 V. cholerae 01 strains isolated in 1997; 98.6%, 93.6%, 83%, 81.9%, 36.2%, 35.5%, 3.2% were sensitive to ciprofloxacin, tetracycline, ampicillin, erythromycin, nalidixic acid, chloramphenicol and trimethoprim/sulphamethoxazole, respectively. Among the 87 V. cholerae 01 isolates collected in 1999, 100%, 58.6%, 46.0%, 46%, 47.1%, 19.5%, 3.4% were sensitive to ciprofloxacin, tetracycline, ampicillin, erythromycin, chloramphenicol, nalidixic acid and trimethoprim/sulphamethoxazole, respectively. Between 1997 and 1999, there was a significant increase in the proportion of V. cholerae 01 isolates resistant to tetracycline, ampicillin, nalidixic acid and to erythromycin but there was no change for susceptibility to ciprofloxacin and trimethoprim/sulphamethoxazole., Conclusion: Significant proportion of V. cholerae 01 strains in Dar es Salaam were resistant to commonly used antimicrobial agents during the two years of the study. Therefore, there is a great need to control the utilisation of antimicrobial agents in cholera control, in addition to continuing carrying out surveillance of antimicrobial resistance as a guide to choice of antimicrobial treatment. Rotational use of the available drugs with regular monitoring of susceptibility may contribute to continuing usefulness of such drugs.

Objective: To determine the antimicrobial susceptibility pattern of S. aureus isolates including the presence of methicillin resistant S. aureus strains., Design: Cross-sectional study., Setting: Department of Microbiology and Immunology, Muhimbili Medical Centre, Dar es Salaam, Tanzania between October 1997 and March 1998., Patients: Two hundred and sixty patients consisting of 67 neonates, 114 children aged 18 years and below and 79 adults., Main Outcome Measures: Antimicrobial susceptibility to tetracycline, erythromycin, cefuroxime, methicillin and penicillin G and presence of mec A gene., Results: Among the S. aureus strains, 97.3%, 68.1%, 37.3% and 6.5% were sensitive to cefuroxime, erythromycin, tetracycline and penicillin G respectively. Only one (0.4%) S. aureus isolate was resistant to methicillin using both the E test and presence of mec A gene. There was no significant difference between the sensitive S. aureus isolates from the neonates, children and adults., Conclusion: S. aureus strains are becoming more resistant to commonly used antimicrobial agents, the prevalence of methicillin resistant S. aureus strains in our study population is low compared with reported studies.

In northwestern Tanzania, a population-based survey of HIV-1 infection in the Kagera region in 1987 demonstrated a high prevalence (24.2%) in adults of Bukoba town, whereas it was lower (10.0%) in the surrounding rural district of Bukoba. In 1993 and 1996, population-based cross-sectional studies were carried out in urban and rural Bukoba districts, respectively, to monitor the time trend in the prevalence of HIV-1 infection in the region. In both studies, a multistage cluster sampling technique was adopted in selecting study individuals. Consenting individuals between 15 and 54 years of age were interviewed using a structured questionnaire. Following individual counseling, blood samples were drawn and tested for HIV infection using enzyme-linked immunosorbent assay (ELISA) antibody detection tests. The overall age-adjusted HIV-1 seroprevalence in urban Bukoba decreased from 24.2% (134 of 553) in 1987 to 18.3% (118 of 653) in 1993 (p = .008). The age-adjusted gender-specific prevalence declined significantly in women, from 29.1% (95 of 325) to 18.7% (74 of 395; p = .0009). Except for men > or = 35 years of age, whose prevalence appeared to have an upward trend between the two studies, all other age groups in both genders had a downward trend; this finding was most significant in women between 15 and 24 years of age (from 27.6% to 11.2%; p = .0004). For the rural population, the overall prevalence decreased from 10.0% (54 of 538) in 1987 to 6.8% (118 of 1728) in 1996 (p = .01). Except for rural women between 15 and 24 years of age whose prevalence decreased from 9.7% (12 of 124) to 3.1% (12 of 383; p = .002), other age groups in the rural populations showed no change in prevalence. Ongoing interventions in this area leading to behavioral change may have contributed to this observation. An incidence study is under way to confirm this observation and to investigate the factors that are responsible for the decline in the HIV-1 prevalence.

Objective: To study late postnatal transmission of human immunodeficiency virus type 1 in a cohort of children born to HIV-1-seropositive mothers who delivered at Muhimbili Medical Centre in Dar es Salaam, Tanzania., Materials and Methods: Since 1991 a prospective cohort study of mother-to-child transmission of HIV-1 has been conducted at Muhimbili Medical Centre in Dar es Salaam. HIV-1-seropositive mothers and age-matched seronegative controls were recruited into the cohort at delivery together with their newborns. Diagnosis of HIV-1 infection in children was based on polymerase chain reaction, HIV-1 p24 antigen tests and HIV antibody tests. Late postnatal transmission was defined as HIV-1 infection occurring after 6 months of age in a child who was uninfected at 6 months of age and who had an HIV-1-seropositive mother. Children born to HIV-seronegative mothers were used as controls. Breast-feeding was universal in this cohort. CD4 and CD8 T lymphocytes were assayed by flow cytometry in the mothers., Results: Among 139 children born to HIV-1-seropositive mothers and known to be HIV-uninfected at 6 months of age, 8 children became HIV-1-infected at the end of their first year of life or later. No conversions were observed in children younger than 11 months. The 8 conversions were observed during a follow-up covering 1555 child months between 6 and 27 months of age corresponding to a conversion rate of 6.2 per 100 child years. Among 260 children with HIV-seronegative mothers no child became HIV-infected during the follow-up. The percentage of CD4 T lymphocytes was similar in mothers with early and late transmission but was significantly lower in transmitting than in nontransmitting mothers., Conclusion: Because no HIV-1 infection occurred in children with HIV-seronegative mothers, we conclude that the observed infections at the end of the first year of life or later among children born to HIV-seropositive women were caused by late transmission from mother to child, most likely through breast-feeding.

In a period of two months, 232 consecutive urinary tract pathogens were isolated from hospitalised and non-hospitalised patients. Among the isolates, 200 (86.2%) were gram negative bacilli, including E. coli 109 (54.5%), Klebsiella species, 44 (22.5%), Enterobacter species 19 (9.5%), Proteus species 18 (9%), Morganella morganii 9 (4.5%) and Salmonella typhimurium, one (0.5%). Antimicrobial susceptibility testing to amoxycillin/clavulanic acid, nitrofurantoin, gentamicin and cefuroxime was performed using Stoke's method. Among the 109 E. coli isolates, 107 (98.2%), 104 (94.5%), 105(95.5% and 107 (98.2%) were sensitive to amoxycillin/clavulanic acid, cefuroxime, nitrofurantoin and gentamicin, respectively. Of the 44 Klebsiella isolates, 42 (95.5%), 41 (95.5%), 40 (90.9%) and 34 (77.3%) were sensitive to amoxycillin/clavulanic acid, cefuroxime, nitrofurantoin and gentamicin, respectively. There was no significant difference when the suceptibility patterns of isolates from hospitalised patients were compared to those from outpatients. Although the susceptibility pattern of urinary tract pathogens to the commonly used antimicrobial agents in the hospital is still favourable, there is a need to establish strategies to prevent emergence of resistant bacterial strains.

Antimicrobial resistance has become a major medical and public health problem. The main factor responsible for development and spread of bacterial resistance is injudicious use of antimicrobial agents which has resulted in most gram positive and gram negative bacteria continuously developing resistance to the antimicrobials in regular use at different time periods. In East Africa, among E. coli in urinary tract infections, more than 80% are currently resistant to ampicillin, cotrimoxazole and tetracycline while more than 80% of the isolates are still susceptible to nitrofurantoin, gentamicin and third generation cephalosporins. Penicillin G resistant strains of pneumococci were first reported in 1967 but had gradually increased to about 20% in 1991. Among group A streptococci, all natural strains are still sensitive to penicillin G while resistance to tetracycline has reached alarming proportions. In Tanzania, more than 65% of N. gonorrhoeae isolates are beta-lactamase producers, however, spectinomycin, second and third generation cephalosporins and ciprofloxacin are effective against most strains. Vibrio cholerae 01 strains resistant to multiple antibiotics are widely spread globally, however, there are recent reports indicating that withdrawal of the drugs can lead to loss of the antibiotic resistance factors. Despite varied susceptibility of N. meningitidis strains world wide, isolates in Tanzania are still susceptible to commonly available drugs including penicillin G and chloramphenicol. Available methods for control of spread of bacterial resistance include rational use of antimicrobial agents including control in animal husbandry, change to newer antimicrobials, rotational use of drugs and constant surveillance for emerging bacterial resistance.

In order to assess the prognostic value of lymphocyte subsets and immune activation markers in HIV-1 infected Tanzanian patients, peripheral white blood cell(WBC) count, total lymphocytes, CD4+ and CD8+ T-lymphocytes and Beta-2 microglobulin (B-2M) concentrations were determined among healthy HIV-1 seronegative Tanzanian blood donors and in infected Tanzania individuals in different clinical stages of HIV-1 infection. CD4+ T-lymphocytes, CD8+ T-lymphocyte percentages, CD4:CD8 lymphocyte ratios and the concentrations of B-2M were strongly correlated with the clinical stages of HIV-1 infection. These results suggest that B-2M could be a useful prognostic marker in HIV-1 infection in settings where T-lymphocyte subset determinations cannot be done.

Objective: To evaluate a WHO testing strategy based on the use of two consecutive enzyme-linked immunosorbent assays (ELISA) as an alternative to ELISA followed by Western blotting (WB) for the serologic diagnosis of HIV infection., Study Design: The study was of 2069 consecutive serum specimens from patients suspected of HIV infection received for HIV diagnostic testing at the HIV laboratory, Muhimbili Medical Centre, Dar es Salaam. The strategy involved testing all sera with Behring indirect anti-HIV 1 + 2 peptide ELISA, followed by Wellcozyme anti-HIV-1 recombinant competitive ELISA on those sera reactive by the first ELISA. WB was done on a sample of the sera reactive on both ELISAs and on all those giving discordant results on the two ELISAs. Of the 2069 sera tested, 837 (40.5%) were negative on the first ELISA, 1172 (56.6%) were reactive on both ELISAs and 60 (2.9%) were initially reactive on the first test but not on the second assay., Results: Of the 1172 sera reactive on both ELISAs, 329 (28.1%) were tested by WB. The diagnostic accuracy of the WHO alternative testing strategy using WB confirmation as the 'gold' standard was as follows: sensitivity 99.4% (326/328), specificity 99.7%, (893/896), positive predictive value 99.1% (328/331) and negative predictive value 99.8% (893/895). Repeated testing by ELISA of the sera which initially gave discordant results on the two ELISAs increased the sensitivity to 100%. Three sera giving false positive reactions on both ELISAs became negative on both ELISAs after retesting. In order to achieve a specificity and a positive predictive value of 100%, it would have been necessary to subject all sera reacting on both ELISAs to retesting on one ELISA., Conclusions: A second ELISA based on different antigens and a different test principle compared with the first ELISA could be used as an alternative to the WB assay for confirmation of HIV antibodies. However, some modifications of the WHO strategy for diagnostic HIV antibody testing were required in order to maximize the diagnostic accuracy.

Alternatives to confirmation of human immunodeficiency virus (HIV)-1 seropositivity by Western blot analysis were evaluated retrospectively using combinations of six anti-HIV-1 screening assays, including four enzyme-linked immunosorbent assays (ELISA) and two simple tests (a rapid dot immunoassay and an agglutination assay), according to an algorithm where sera are first screened by one assay and those repeatedly reactive on this assay are tested repeatedly by a second assay. Two panels of sera collected in Dar es Salaam, Tanzania, were used. Panel 1 was composed of 1,465 consecutive blood donor sera of which 99 (6.8%) were confirmed HIV-1 antibody positive, and panel 2 was composed of sera from 396 consecutively admitted patients at two medical wards of which 116 (29.3%) were confirmed HIV-1 antibody positive. Sera reactive on any of the six screening assays were also tested by a confirmatory Western blot assay. The sensitivity of the assays at the initial valid testing were as follows: Abbott 99.5%, Behring 99.5%, Organon 97.7%, Wellcozyme 100%, HIV CHEK-1 95.8%, and Serodia 95.8%. After repeat testing of sera that initially gave false-negative results all assays showed 100% sensitivity except HIV CHEK-1 (98.6%). The specificities after repeat testing were between 99.6 and 99.9% for all assays except for the Behring ELISA (98.1%). Several combinations of screening assays were found to give the same diagnostic accuracy as the screening assay followed by Western blot analysis. We conclude that an alternative confirmatory strategy can be fully satisfactory for some testing purposes.