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INTRODUCTION The prevalence of azole resistance in C parapsilosis is very low in most parts of the world. However, South Africa has reported an exceptionally high prevalence of azole resistance in C parapsilosis strains isolated from candidaemia cases. We aimed to determine the possible molecular mechanisms of fluconazole resistance in C parapsilosis isolates obtained through surveillance at a large neonatal unit at a South African academic hospital. METHODS We sequenced the ERG11 and MRR1 genes of C parapsilosis isolates recovered from cases of neonatal candidemia, followed by microsatellite genotyping. A total of 73 isolates with antifungal susceptibility results were analysed. RESULTS Of these, 57 (78%) were resistant, 11 (15%) susceptible dose-dependent and 5 (7%) susceptible. The most commonly identified amino acid substitution within the ERG11 gene was Y132F in 68% (39/57) of fluconazole-resistant isolates and none in susceptible isolates. Three amino acid substitutions (R405K, G583R and A619V) and 1 nucleotide deletion at position 1331 were identified within MRR1 gene in 19 (26%) isolates. Microsatellite genotyping grouped isolates into four clusters (50 isolates). Cluster 1 accounted for 23% (17/73) of all cases, cluster 2 for 22% (16/73), cluster 3 for 14% (10/73) and cluster 4 for 10% (7/73). We found an association between cluster type and fluconazole resistance (P-value = .004). Isolates harbouring the Y132F substitution were more likely to belong to a cluster than non-Y132F isolates. CONCLUSION Fluconazole resistance in C parapsilosis strains from a single South African neonatal unit was associated with cluster type and predominantly driven by Y123F amino acid substitutions in the ERG11 gene.

Purpose: The present study was done to evaluate the antimycobacterial activity of aqueous and methanol extracts of nine plants viz., Buddleja saligna, Carpobrotus dimidiatus, Capparis tomentosa, Dichrostachys cinerea, Ekebergia capensis, Ficus sur, Gunnera perpensa, Leonotis leonurus and Tetradenia riparia in South Africa. Methods: Aqueous and methanol extracts of the leaves the plants were tested in vitro for their activity against Mycobacterium smegmatis, Mycobacterium tuberculosis H37Rv (ATCC 25177) and three well-characterized clinical isolates of MDR and XDR-TB isolates using the agar incorporation method. The minimum inhibitory concentration (MIC) of each of the active plant extract was determined using the broth microdilution method. Cytotoxic effect was evaluated against the mouse BALB/C monocyte macrophage (J774.2) and peripheral blood mononuclear cells (PBMCs) whole the toxicity screening was done using the brine shrimp lethality assay. Composition of each of the plants was determined using thin layer chromatography while qualitative analysis of antimycobacterial compounds was done using TLC-Bioautography Results: The methanol extracts of B. saligna, C. tormentosa and C. dimidiatus; and aqueous extracts of G. perpensa and T. riparia possessed significant activity against M. smegmatis, M. tuberculosis H37Rv (ATCC 25177) and the three well-characterized clinical isolates of MDR and XDR-TB. Except for a high concentration of G. perpensa none of the other plants which possessed antimycobacterial activity showed any toxic or cytotoxic activity. Conclusion: Our findings show that B. saligna, C. tormentosa, C. dimidiatus, G. perpensa, and T. riparia have antimycobacterial activity. Further studies would aim at isolation and identification of the active compounds from the plants extracts which had positive antimycobacterial activity.

Neonatal candidaemia is a common, deadly and costly hospital-associated disease. To determine the genetic diversity of Candida parapsilosis causing fungaemia in South African neonatal intensive care units (NICUs). From February 2009 through to August 2010, cases of candidaemia were reported through laboratory-based surveillance. C. parapsilosis isolates from neonatal cases were submitted for identification by internal transcribed spacer (ITS) region sequencing, antifungal susceptibility testing and microsatellite genotyping. Cluster analysis was performed using Unweighted Pair Group Method with Arithmetic Mean (UPGMA). Of 1671 cases with a viable Candida isolate, 393 (24%) occurred among neonates. Isolates from 143 neonatal cases were confirmed as C. parapsilosis sensu stricto. Many isolates were resistant to fluconazole (77/143; 54%) and voriconazole (20/143; 14%). Of 79 closely-related genotypes, 18 were represented by ≥2 isolates; 61 genotypes had a single isolate each. Seven clusters, comprised of 82 isolates, were identified at five hospitals in three provinces. Isolates belonging to certain clusters were significantly more likely to be fluconazole resistant: all cluster 7 isolates and the majority of cluster 4 (78%), 5 (89%) and 6 (67%) isolates (P

BACKGROUND Capreomycin is a key antimycobacterial drug in treatment of extensively drug-resistant tuberculosis (XDR-TB). Drug-susceptibility testing (DST) for capreomycin is not routinely performed in newly diagnosed XDR-TB in South Africa. We performed this study to assess the prevalence, clinical significance, and molecular epidemiology of capreomycin resistance in newly diagnosed patients with XDR-TB in KwaZulu-Natal, South Africa. METHODS Retrospective cohort study of consecutive patients with XDR-TB admitted to a TB referral hospital without previous XDR-TB treatment. A subset of isolates had extended DST (including capreomycin), mutational analysis, and IS6110 restriction fragment length polymorphism assays. RESULTS A total of 216 eligible patients with XDR-TB were identified. The majority were treated with capreomycin (72%), were young (median age: 35.5 years), and were female (56%). One hundred five (76%) were HIV+, and 109 (66%) were on antiretroviral therapy. A subset of 52 patients had full DST. A total of 47/52 (90.4%) patients with XDR-TB were capreomycin resistant. Capreomycin-resistant patients experienced worse mortality and culture conversion than capreomycin susceptible, although this difference was not statistically significant. The A1401G mutation in the rrs gene was associated with capreomycin resistance. The majority of capreomycin-resistant strains were F15/LAM4/KZN lineage (80%), and clustering was common in these isolates (92.5%). CONCLUSIONS Capreomycin resistance is common in patients with XDR-TB in KwaZulu-Natal, is predominantly because of ongoing province-wide transmission of a highly resistant strain, and is associated with high mortality. Capreomycin should be included in routine DST in all patients with XDR-TB. New drug regimens that do not include injectable agents should be operationally tested as empiric treatment in XDR-TB.

We determined the susceptibility of 102 clinical isolates Cryptococcus neoformans from Durban, South Africa, to amphotericin B, fluconazole, flucytosine, and voriconazole using broth microdilution (BMD) according to the Clinical and Laboratory Standards Institute M27-A3 document and compared these results with Etest and Vitek 2 ® . Essential agreement (EA) of Etest and Vitek 2 ® compared to BMD was determined. Low MICs that were below the epidemiological cutoff values of the 4 antifungal agents tested were demonstrated by all isolates. The EA of Etests for fluconazole, amphotericin, and voriconazole was 95.1%, 83.3%, and 91.2%, respectively, and for Vitek 2 ® EA for fluconazole, amphotericin, and flucytosine was 97.1%, 95.1%, and 97.1%, respectively. The Vitek 2 ® showed good agreement with BMD and is a suitable alternative. Etests demonstrated good EA for azoles only. Clinical cryptococcal isolates from Durban remain susceptible to current recommended antifungal therapy.

Background: Hospital acquired infections are on the increase worldwide. A possible source for transmission is the presence of microorganisms on mobile phones which are carried by increasing numbers of medical and nursing staff, students, and caregivers.Methods: A cross-sectional study was performed. Samples were obtained from medical and nursing staff, students, and caregivers from the paediatric wards (85 beds), and the neonatal unit (40 beds) at King Edward VIII Hospital (KEH), Durban. Mobile phones of participants were sampled, with sterile swabs, without prior warning. The swabs were transported promptly to the laboratory and cultured onto colistin, nalidixic acid agar and MacConkey agar plates. All positive cultures were identified using standard laboratory tests.Results: Of the 100 mobile phones sampled, 30 were contaminated with bacteria. Gram-positive microorganisms were more frequently cultured than Gram-negative microorganisms (29 vs. 7). Significantly more caregivers had contaminated phones (54.17% vs. 22.37%, p value = 0.003). Caregivers’ phones were predominantly contaminated with Staphylococcus spp. (41.67%). More phones in the general wards (37.5%) and nursery (32.5%) were contaminated compared to the gastroenteritis ward (10%).Conclusions: Our results indicate that one third of the mobile phones carried in the paediatric wards are contaminated. Caregivers had the highest likelihood of carrying mobile phones that were contaminated, with the lowest rate among the nursing staff. Caregivers’ phones were also more likely to have multiple microorganisms. The lowest rate of contamination was found in the gastroenteritis ward, possibly due to a heightened awareness of the need for handwashing.

Buddleja saligna (family Buddlejaceae) is a medicinal plant endemic to South Africa. Two isomeric pentacyclic triterpenes, oleanolic acid and ursolic acid, were isolated from the leaves of B. saligna using silica gel column chromatography. Compounds oleanolic acid and ursolic acid were subjected to derivatization with acetic anhydride in the presence of pyridine to obtain oleanolic acid-3-acetate and ursolic acid-3-acetate, respectively. The structures of these compounds were fully characterized by detailed nuclear magnetic resonance (NMR) investigations, which included

Streptococcus agalactiae is a well-established cause of neonatal meningitis, whereas Acinetobacter baumannii, often seen as a contaminant, has only rarely been reported to cause meningitis without a neurosurgical predisposition. We report of a case of neonatal meningitis in a preterm neonate in which both S. agalactiae and A. baumannii were considered the pathogens. This case highlights the pathogenicity of A. baumannii, and suggests that it may be frequently underestimated.

Background: Cryptococcal meningitis is the most common systemic fungal infection in HIV infected patients. The India ink stain is currently recommended for the microscopic detection of cryptococci. There is very little published data on the utility of the Gram stain. We therefore, undertook a comparative evaluation of the Gram stain and India ink stain for the rapid diagnosis of cryptococcal meningitis in HIV infected patients.Methodology: Gram and India ink stains were performed on 90 cerebrospinal fluid (CSF) samples. Culture was used as the gold standard.Results: Of the 83 culture positive samples, the India ink was positive in 84% [70 /83] and the Gram stain was positive in 89% [74/83]. The specificity for both methods was 100%.Discussion: Our results indicate that the Gram stain is a simple, cheap and reliable method for the diagnosis of cryptococcal meninigitis. It is routinely performed on all CSF samples received in the laboratory and would therefore detect cryptococci in cases where the c...

Objective HIV-infected patients with treated cryptococcal meningitis are at risk for further neurological deterioration after commencing combination antiretroviral therapy (cART), mostly because of cryptococcosis-associated immune reconstitution inflammatory syndrome (C-IRIS). Identifying predictors of C-IRIS could enable risk stratification. Design Prospective, longitudinal cohort study for 24 weeks. Setting Durban, South Africa. Participants One hundred and thirty HIV-infected patients with first cryptococcal meningitis episode Intervention : Antifungal therapy (amphotericin 1 mg/kg median 14 days, followed by consolidation and maintenance fluconazole) and cART (commenced median of 18 days from cryptococcal meningitis diagnosis). Main outcome measure Clinical, blood, and cerebrospinal fluid (CSF) markers associated with C-IRIS before and during cART and clinical significance of CSF cryptococcal culture negativity pre-cART commencement. Results Of 106 patients commencing cART, 27 (25.5%) developed C-IRIS, 16 (15.1%) neurological deterioration-not C-IRIS, and 63 (59.4%) no neurological deterioration. On multivariable analysis, C-IRIS was associated with persistent CSF cryptococcal growth [hazard ratio (HR) 0.27, P=0.026] and lower CSF protein (HR 0.53, P=0.059) prior to cART and lower CD4 T-cell increases (HR 0.99, P=0.026) but not change in HIV viral load during cART. Using survival analysis, patients with a negative cryptococcal culture pre-cART commencement (n=51; 48.1%) experienced fewer episodes of neurological deterioration, C-IRIS, and cryptococcal relapse/persistence than patients with culture positivity (n=55; 51.9%, HR 0.33, 0.33, and 0.12 and P=0.0003, 0.0042, and 0.0004, respectively). Conclusion Persistent CSF cryptococcal growth at cART initiation and poor CD4 T-cell increases on cART are strong predictors of C-IRIS. Approaches aimed at achieving CSF culture negativity prior to cART should be evaluated as a strategy to reduce rates of C-IRIS.

Background: Anaesthetic equipment is a potential vector for the transmission of disease. This study was undertaken to observe current infection control practices among anaesthetic nurses regarding the decontamination of anaesthetic equipment in regional, tertiary and central hospitals in KwaZulu-Natal.Method: All hospitals that were classified as regional, tertiary and central hospitals on the KwaZulu-Natal Department of Health website (15 in total) were visited. All available anaesthesia nurses were invited to participate in a structured interview.Results: Thirty-four anaesthesia nurses were interviewed. Results revealed that decontamination of anaesthetic equipment and other infection control practices were inadequate or inappropriate in several of the hospitals. Practices varied from one healthcare facility to another, as well as within the same facility.Conclusion: Current infection control practices among anaesthesia nurses regarding the decontamination of anaesthetic equipment in the observed hospitals are poor. In light of the high prevalence of many infectious diseases, in particular human immunodeficiency virus, hepatitis B and tuberculosis in KwaZulu-Natal, it is critical that issues relating to decontamination practices are urgently addressed.Keywords: anaesthetic equipment, contamination, infection control, anaesthesia

From February 2009 through August 2010, we compared species-level identification of bloodstream Candida isolates and susceptibility to fluconazole, voriconazole, and caspofungin between diagnostic and reference South African laboratories during national surveillance for candidemia. Diagnostic laboratories identified isolates to genus/species level and performed antifungal susceptibility testing, as indicated. At a reference laboratory, viable Candida isolates were identified to species-level using automated systems, biochemical tests, or DNA sequencing; broth dilution susceptibility testing was performed. Categorical agreement (CA) was calculated for susceptibility results of isolates with concordant species identification. Overall, 2172 incident cases were detected, 773 (36%) by surveillance audit. The Vitek 2 YST system (bioMerieux Inc, Marcy l'Etoile, France) was used for identification (360/863, 42%) and susceptibility testing (198/473, 42%) of a large proportion of isolates. For the five most common species (n = 1181), species-level identification was identical in the majority of cases (Candida albicans: 98% (507/517); Candida parapsilosis: 92% (450/488); Candida glabrata: 89% (89/100); Candida tropicalis: 91% (49/54), and Candida krusei: 86% (19/22)). However, diagnostic laboratories were significantly less likely to correctly identify Candida species other than C. albicans versus C. albicans (607/664, 91% vs. 507/517, 98%; P < .001). Susceptibility data were compared for isolates belonging to the five most common species and fluconazole, voriconazole, and caspofungin in 860, 580, and 99 cases, respectively. Diagnostic laboratories significantly under-reported fluconazole resistance in C. parapsilosis (225/393, 57% vs. 239/393, 61%; P < .001) but over-reported fluconazole non-susceptibility in C. albicans (36/362, 10% vs. 3/362, 0.8%; P < .001). Diagnostic laboratories were less likely to correctly identify Candida species other than C. albicans, under-reported fluconazole resistance for C. parapsilosis and over-reported fluconazole resistance for C. albicans.

A series of 2-(substituted phenyl/benzyl-amino)-6-(4-chlorophenyl)-5-(methoxycarbonyl)-4-methyl-3,6-dihydropyrimidin-1-ium chlorides 7-13 and 15 was synthesized in their hydrochloride salt form. The title compounds were characterized by FT-IR, NMR ((1)H and (13)C) and elemental analysis. They were evaluated for their in vitro antitubercular activity against Mycobacterium tuberculosis H37Rv, multidrug resistance tuberculosis and extensively drug resistance tuberculosis by agar diffusion method and tested for the cytotoxic action on peripheral blood mononuclear cells by MTT assay. Among all the tested compounds in the series, compounds 7 and 11 emerged as promising antitubercular agents at 16 μg/mL against multidrug resistance tuberculosis and over 64 μg/mL against extensively drug resistance tuberculosis. The conformational features and supramolecular assembly of the promising compounds 7 and 11 were determined by single crystal X-ray study.

We conducted a study to determine the frequency of bacteraemias following dental extraction and common oral procedures, namely tooth brushing and chewing, and the relationship between bacteraemia and oral health in black patients. Positive blood cultures were detected in 29.6% of patients after dental extraction, in 10.8% of patients after tooth brushing and in no patients after chewing. No relationship between the state of oral health, which was assessed using the plaque and gingival indices, and the incidence of bacteraemia was found. The duration of bacteraemia was less than 15 minutes. One patient had a positive blood culture prior to dental extraction; his oral health status was poor. Our study confirmed that bacteraemia occurs after tooth brushing.

The antifungal and antimicrobial activities of three pentacycloundecane (PCU) tetra-amine derivatives are reported herein. The in vitro activity of these PCU derivatives against yeasts (Candida albicans and non-albicans species) and filamentous fungi was evaluated using the Clinical and Laboratory Standards Institute (CLSI) M27-A2 and M38-A2 guidelines and the 2H-tetrazolium salt, (MTS) colorimetric method. The minimum inhibitory concentration against most of the tested clinical fungal strains for GKM8 and GKM9 derivatives ranges from 15.6 to 62.5 μg/mL while GKM11 ranged from 3.9 to 7.8 μg/mL. The GKM11 derivative was also active against fluconazole-resistant strains of fungi. The GKM11 derivative also exhibited promising activity against filamentous fungi in that it was 2.5 times more active than amphotericin B against Sporothrix schenckii. Antibacterial activity was determined using the broth microdilution method (BMM) and the iodonitrotetrazolium chloride (INT) colorimetric method. The GKM11 derivative was mainly active against Gram-positive bacteria with MIC ranging from 3.9 to 7.8 μg/mL. Activity against Gram-negative bacteria tested was limited to Escherichia coli and Elizabethkingia meningoseptica (MIC of 31 μg/mL).

The antimicrobial activity of five novel SQ109 derivatives including SQ109 against bacteria, yeast and filamentous fungi is reported herein. Using broth microdilution techniques, compounds 2 and 3 were found to be active against most tested fungi and bacteria, with minimum inhibitory concentrations (MICs) ranging from 0.98 to 31 μg ml−1, except for Klebsiella pneumonia where the MIC was 250 μg ml−1. SQ109 and derivative 4 did not show any significant activity against most of the organisms used. However, their reduced derivatives 1 and 5 showed promising activity with MICs between 0.49 and 62.5 μg ml−1 against most of the microorganisms used.

Current tools for the rapid diagnosis of tuberculous meningitis (TBM) are suboptimal. We evaluated the clinical utility of a quantitative RD-1 IFN-gamma T-cell enzyme-linked immunospot (ELISPOT) assay (T-SPOT.TB), using cerebrospinal fluid cells for the rapid immunodiagnosis of TBM.To evaluate the diagnostic utility of the RD1 antigen- specific ELISPOT assay for the diagnosis of tuberculous meningitis.The ELISPOT assay was evaluated in 150 patients with suspected TBM who were categorized as definite-TBM, probable-TBM, and non-TBM. Culture or polymerase chain reaction positivity for Mycobacerium tuberculosis served as the reference standard. To determine the diagnostic value of the ELISPOT assay, a clinical prediction rule was derived from baseline clinical and laboratory parameters using a multivariable regression model.A total of 140 patients (81% HIV-infected; median CD4 count, 160 cells/mm(3)) were included in the final analysis. When comparing the definite-TBM (n = 38) and non-TBM groups (n = 48), the ELISPOT assay (cut point ofor =228 spot-forming cells per 1 million mononuclear cells) was a useful rule-in test: sensitivity 58% (95% confidence interval [CI], 41-74); specificity 94% (95% CI, 83-99). However, ELISPOT outcomes improved when other rapid tests were concurrently used to exclude bacterial (Gram stain) and cryptococcal meningitis (latex-agglutination test) within the non-TBM group. Using this approach, the ELISPOT assay (cut point ofor =46 spot-forming cells) was an excellent rule-in test: sensitivity 82% (95% CI, 66-92); specificity 100% (95% CI, 78-100); positive predictive value, 100% (95% CI, 89-100); negative predictive value, 68% (95% CI, 45-86); area under the curve, 0.90. The ELISPOT assay had incremental diagnostic value compared with the clinical prediction rule.The RD-1 ELISPOT assay, using cerebrospinal fluid mononuclear cells and in conjunction with other rapid confirmatory tests (Gram stain and cryptococcal latex-agglutination test), is an accurate rapid rule-in test for TBM in a TB and HIV endemic setting.

OBJECTIVES To compare Candida species distribution and antifungal susceptibility at South African public- and private-sector hospitals. METHODS From February 2009 through to August 2010, laboratory-based surveillance for candidaemia was undertaken at 11 public-sector hospitals and >85 private-sector hospitals across South Africa. A case was defined as a patient of any age admitted to a sentinel hospital with isolation of Candida species from blood culture. Viable isolates were identified and tested for antifungal susceptibility at a reference laboratory. Demographic and limited clinical data were abstracted from laboratory records. RESULTS In total, 2172 cases of candidaemia were detected. Among patients with available data, almost two-thirds were critically ill (719/1138, 63%). On multivariable analysis, neonates [adjusted OR (aOR), 2.2; 95% CI, 1.5-3.1; P

Objectives: Unsafe injection and vial usage practices, including the reuse of needles and syringes for different patients, is one of the leading causes of the iatrogenic spread of blood-borne diseases. A study was conducted to determine the prevalence of the reuse of single-patient syringes and spinal fentanyl ampoules among anaesthetists at regional, tertiary and central hospitals in KwaZulu-Natal.Method: All hospitals that are classified as regional, tertiary and central hospitals on the KwaZulu-Natal Department of Health website were visited. All encountered anaesthetists, regardless of rank or experience, were invited to complete a simple questionnaire in confidence.Results: Ninety-one anaesthesiologists and anaesthetic practitioners completed the questionnaire. Thirteen (14%) of the anaesthetists admitted to reusing syringes on different patients. Seventeen (19%) of the anaesthetists admitted to reusing syringes on different patients after they had changed the needle or set. Fifty-seven (63%)...

To create a prognostic model for complications of blood culture-proven typhoid fever in adults (> or = 15 y old), a retrospective cohort was assembled though review of the medical records of the hospitalized patients treated for typhoid fever over a 3-y period ending January 1995. Of the 59 patients included, 21 (35.6%) developed various complications of typhoid fever. No patient included died. Four baseline variables (abdominal pain, systolic blood pressure < 100 mmHg, hypoalbuminaemia < 32 g/l and laboratory evidence of disseminated intravascular coagulation) were independently associated with complications and were used to create a prognostic model. The prediction accuracy of the model was determined using the concordance index (c-index). The results (c-index, 0.915 [95%, CI, 89.0-93.0]) showed that the model predicted complications significantly better than chance. The model stratified patients into 3 prognostic stages: low risk for complications (0%; stage I), intermediate risk (42.9%; stage II) and high risk (92.3%; stage III) (p = 0.001). If validated in other settings, it will help clinicians in predicting complications in patients with blood culture-proven typhoid fever on admission.

Clinical features, laboratory findings, and complications of typhoid fever were correlated with sex through a retrospective case note review of 102 hospitalized culture-positive patients in Durban, South Africa. Intestinal perforation (P = 0.04), occult blood losses in stools (P = 0.04), and a mild reticulocytosis in the absence of hemolysis (P = 0.02) occurred more frequently in males than in females. A single pretreatment Widal O antibody titer > or = 1:640 was also a statistically significant occurrence in males (P = 0. 006). Female patients were significantly more severely ill (P = 0.0004) on admission and had chest signs consistent with bronchopneumonia (P = 0.04), transverse myelitis (P = 0.04), abnormal liver function test results (P = 0.0003), and abnormal findings in urinalyses (P = 0.02). Typhoid hepatitis (P = 0.04) and glomerulonephritis (P = 0.02) were present significantly more frequently in females. Whether these differences were due to differences in host's immune response to acute infection need to be determined in a prospective study.

Ten patients with asymptomatic human immunodeficiency virus (HIV) infection were treated for typhoid fever at King Edward VIII Hospital, Durban, South Africa, from 1993 through 1995. The mean age was 23.7 years (range, 8-33 years), with a female-to-male ratio of 9 to 1 and mortality and morbidity rates of 20% and 10%, respectively. Common presenting manifestations were fever (100%), relative bradycardia (50%), and diarrhea (40%). With respect to epidemiologic and clinical characteristics, we noted no significant differences among these 10 HIV-positive and 32 HIV-negative patients treated for typhoid fever during the same period. However, we found hepatic dysfunction in the form of an isolated increase in aspartate aminotransferase (p < 0.01) and abnormal urinary findings suggestive of glomerulonephritis (p = 0.01) more frequently in HIV-positive patients.

SETTING: The dual epidemics of HIV-TB including MDR-TB are major contributors to high morbidity and mortality rates in South Africa. Rifampicin (RIF) resistance is regarded as a proxy for MDR-TB. Currently available molecular assays have the advantage of rapidly detecting resistant strains of MTB, but the GeneXpert does not detect isoniazid (INH) resistance and the GenoTypeMTBDRplus(LPA) assay may underestimate resistance to INH. Increasing proportions of rifampicin mono-resistance resistance (RMR) have recently been reported from South Africa and other countries. OBJECTIVE: This laboratory based study was conducted at NHLS TB Laboratory, Durban, which is the reference laboratory for culture and susceptibility testing in KwaZulu-Natal. We retrospectively determined, for the period 2007 to 2009, the proportion of RMR amongst Mycobacterium tuberculosis (MTB) isolates, that were tested for both RIF and INH, using the gold standard of culture based phenotypic drug susceptibility testing (DST). Gender and age were also analysed to identify possible risk factors for RMR. DESIGN: MTB culture positive sputum samples from 16,748 patients were analysed for susceptibility to RIF and INH during the period 2007 to 2009. RMR was defined as MTB resistant to RIF and susceptible to INH. For the purposes of this study, only the first specimen from each patient was included in the analysis. RESULTS: RMR was observed throughout the study period. The proportion of RMR varied from a low of 7.3% to a high of 10.0% [overall 8.8%]. Overall, males had a 42% increased odds of being RMR as compared to females. In comparison to the 50 plus age group, RMR was 37% more likely to occur in the 25-29 year age category. CONCLUSION: We report higher proportions of RMR ranging from 7.3% to 10% [overall 8.8%] than previously reported in the literature. To avoid misclassification of RMR, detected by the GeneXpert, as MDR-TB, culture based phenotypic DST must be performed on a second specimen, as recommended by the SA NDOH TB guidelines as well as WHO. We suggest that two sputum samples should be obtained at the first visit. The second sputum sample should be stored at 4°C. The latter sample is then readily available for performing additional DST (phenotypic or genotypic) for 2nd lines drugs, resulting in a decreased waiting period for DST results to become available.

Neisseria meningitidis is a leading cause of pyogenic meningitis worldwide, as well as causing large epidemics in parts of Africa. With the dramatic decline in cases of Haemophilus influenzae B, N. meningitidis has emerged as one of the most common causes of acute bacterial meningitis in children and adults in South Africa. However, it remains an uncommon cause of meningitis in the neonatal period. Subdural empyema together with hydrocephalus has been infrequently described as a complication of meningococcal meningitis. We report a rare case of neonatal meningococcal meningitis, complicated by subdural empyema and hydrocephalus. To the best of our knowledge only a few cases of neonatal meningococcal meningitis have been reported from South Africa, with none of these cases having the complication of subdural empyema.

We evaluated some of the regimens recommended for the antimicrobial prophylaxis of infective endocarditis prior to dental extraction in 160 patients. Group A patients served as the control group, group B subjects rinsed their mouths with chlorhexidene, group C subjects took 3 g amoxicillin orally and group D patients took 600 mg clindamycin orally. The proportion of patients who had post-extraction bacteraemia in groups A, B, C and D was 35, 40, 7.5 and 20%, respectively. The differences between the control and amoxicillin groups (p = 0.003) and between the chlorhexidine and amoxicillin groups (p = 0.0006) were statistically significant. Streptococci were not isolated in any patients in the amoxicillin and clindamycin groups. In our study, none of the regimens were effective in preventing post-extraction bacteraemia.

Background. Non-tuberculous mycobacteria (NTM) are important environmental pathogens capable of causing a spectrum of infection. The different species exhibit varied geographical prevalence worldwide. Identification of the infecting organism may be helpful in determining the clinical significance of the isolate. Objective. To describe the spectrum of NTM isolated from clinical specimens received at the National Health Laboratory Service central tuberculosis laboratory in KwaZulu-Natal Province, South Africa. Method. In a laboratory-based prospective study, 200 suspected NTM were randomly selected over a period of 1 year and identified to species level using a commercially available DNA strip assay (GenoType Mycobacterium, CM/AS; Hain Lifescience, Germany). Results. Of the 200 suspected NTM, 133 (66.5%) were confirmed to be NTM by the molecular test. The most frequently isolated NTM species were Mycobacterium intracellulare (45.9%), M. avium subspecies (11.3%), M. gordonae (6.0%) and M. kansasii (4.5%). Conclusion. It is important for laboratories to document the local spectrum of NTM because of the geographical variation in the different NTM species isolated. Although molecular tests for identifying NTM are relatively expensive, they have the advantage of providing rapid and accurate identification of the various NTM species.

A retrospective case note review was undertaken to assess the clinical signi® cance of hepatic dysfunction with jaundice in typhoid fever. Of the 57 patients, 21 (36.8% ) had jaundice, while 36 (63.2% ) did not have jaundice. Signi® cantly higher proportions of jaundiced patients were females (P 5 0.04). Confusion (P 5 0.01), upper abdominal pain (P 5 0.02), right upper quadrant tenderness (P 5 0.0001), and low prothrombin index (P 5 0.04) were statistically signi® cant occurrences in jaundiced patients on admission. Admission mean values of serum bilirubin (P 5 0.0001), g -glutamyltranspe ptidase (GGT; P 5 0.009), and alanine aminotransferase (ALT; P 5 0.0005) were signi® cantly higher in icteric patients while mean values of total serum protein (P 5 0.0009) and albumin (P 5 0.0001) were signi® cantly higher in anicteric patients. There were no deaths. Glomerulonephritis occurred signi® cantly (P 5 0.001) more frequently in icteric patients. It is concluded that hepatic dysfunction with jaundice in typhoid fever indicates more severe hepatic injury, which may precipitate the development of clinically detectable glomerulonephritis.

Four cases of typhoid fever complicated by both acute oliguric renal failure and hepatitis are presented. Two patients had type II hepatitis according to criteria proposed by Khosla et al. (30) with hepatomegaly, hyperbilirubinaemia and markedly elevated asparate transaminase (AST); the others had type III hepatitis, characterized clinically and biochemically by profound jaundice, hepatomegaly, hepatic encephalopathy (one case only), hyperbilirubinemia and markedly elevated serum AST. Renal biopsy was not performed in any of our patients. However, a combination of proteinuria and abnormal urinary sediments containing red cell casts and granular casts, as noted in these patients, is considered highly suggestive of glomerulonephritis. Although isolated renal failure and hepatitis with hepatomegaly and deranged liver enzyme values have been reported previously in typhoid fever, their occurrence simultaneously in the same patient in distinctly rare, having been reported only twice in the English language literature.

Vinod Patel and colleagues evaluate the sensitivity and specificity of quantitative PCR using Xpert MTB/RIF for diagnosis of TB meningitis in the high-burden setting of South Africa. Please see later in the article for the Editors' Summary, Background Tuberculous meningitis (TBM) is difficult to diagnose promptly. The utility of the Xpert MTB/RIF test for the diagnosis of TBM remains unclear, and the effect of host- and sample-related factors on test performance is unknown. This study sought to evaluate the sensitivity and specificity of Xpert MTB/RIF for the diagnosis of TBM. Methods and Findings 235 South-African patients with a meningeal-like illness were categorised as having definite (culture or Amplicor PCR positive), probable (anti-TBM treatment initiated but microbiological confirmation lacking), or non-TBM. Xpert MTB/RIF accuracy was evaluated using 1 ml of uncentrifuged and, when available, 3 ml of centrifuged cerebrospinal fluid (CSF). To evaluate the incremental value of MTB/RIF over a clinically based diagnosis, test accuracy was compared to a clinical score (CS) derived using basic clinical and laboratory information. Of 204 evaluable patients (of whom 87% were HIV-infected), 59 had definite TBM, 64 probable TBM, and 81 non-TBM. Overall sensitivity and specificity (95% CI) were 62% (48%–75%) and 95% (87%–99%), respectively. The sensitivity of Xpert MTB/RIF was significantly better than that of smear microscopy (62% versus 12%; p = 0.001) and significantly better than that of the CS (62% versus 30%; p = 0.001; C statistic 85% [79%–92%]). Xpert MTB/RIF sensitivity was higher when centrifuged versus uncentrifuged samples were used (82% [62%–94%] versus 47% [31%–61%]; p = 0.004). The combination of CS and Xpert MTB/RIF (Xpert MTB/RIF performed if CS, Editors' Summary Background Worldwide, tuberculosis (TB) is the leading cause of death among people living with HIV. The risk of developing TB is estimated to be 12–20 times greater in people with HIV than in people without HIV. The World Health Organization reported that, in 2011, there were 8.7 million new cases of TB, of which 1.1 million were among people living with HIV. TB infection in people living with HIV is a major problem in sub-Saharan Africa, where up to 80% of individuals infected with TB are also infected with HIV. TB is caused by a bacterial infection spread through the air from one person to another when the infected person coughs or sneezes, for example. It usually affects the lungs, but it can also affect other parts of the body including the brain, where it leads to meningitis. People with meningitis caused by TB are often seriously ill. Many may develop brain damage, and 30% will die, particularly if they aren't diagnosed quickly and treatment is delayed. TB meningitis is therefore a serious health concern in countries with high rates of HIV and TB co-infection. Why Was This Study Done? There is currently no simple test to diagnose TB meningitis. The tests that are available detect only about 50% of cases. They are expensive and practical to use only in a high-tech environment, and are therefore unsuitable for low-income countries. Recently, a new test has become available to detect TB, known as Xpert MTB/RIF. The test is used to detect the DNA (the molecular biological instructions for each organism) of the bacteria that causes TB. It is accurate at detecting TB lung infection, requires minimal training to operate, and is relatively inexpensive. It is now being used to diagnose TB in countries with high rates of the disease, including South Africa. However, thus far its use has been limited to detecting the TB bacterium in sputum samples (a mixture of saliva and phlegm) from people with a lung infection. The few studies that have assessed whether the test can be used to detect TB meningitis have been small and inconclusive. This study was carried out to determine whether Xpert MTB/RIF could be used to detect TB bacteria in the cerebrospinal fluid (the fluid that surrounds the brain and spinal cord) in people with TB meningitis. The researchers wanted to find out whether the test would be sensitive (correctly identifying patients with TB meningitis) and specific (correctly identifying patients without TB meningitis). They also wanted to address more practical questions such as how much cerebrospinal fluid needs to be collected and how the sample needs to be processed to ensure accurate results. What Did the Researchers Do and Find? The researchers used the Xpert MTB/RIF test to analyze cerebrospinal fluid samples from 204 patients with suspected TB meningitis. These patients were recruited from hospitals in South Africa between January 2008 and December 2011. Standard diagnostic tests were used to categorize these patients as either definitely having TB meningitis, possibly having TB meningitis, or not having TB meningitis. Among patients infected with HIV, the Xpert MTB/RIF correctly identified 62% of those with TB meningitis and 95% of those without TB meningitis. The researchers also assessed whether it would be more cost-effective to use the test only for cases where the standard diagnostic procedure was uncertain, i.e., to avoid testing in cases where TB meningitis was very likely following the normal clinical assessment. Based on the researchers' theoretical analysis, this would reduce test use by only about 10%. What Do These Findings Mean? This study suggests that Xpert MTB/RIF is a useful diagnostic test for TB meningitis in patients infected with HIV living in areas where there are high levels of TB infection. It is not known how well the test would perform in places where TB levels are low, and the test did not perform well in individuals without HIV, although there were very few of these patients. The Xpert MTB/RIF test correctly identified more positive cases than the other tests used to diagnose TB meningitis, within 24 hours of first seeing a patient. However, the test accuracy was best when the cerebrospinal fluid sample was centrifuged (spinning the sample very fast to concentrate the test material) to achieve the best results. This means additional apparatus would be required, resulting in higher cost and requiring more training. The researchers conclude that this test could still be useful in settings where resources are limited. There are also important questions that remain unanswered. This study shows only that the Xpert MTB/RIF test is useful in determining that a patient has TB meningitis. It is not useful in determining that a patient does not have TB meningitis. Further research is needed to determine whether the test will be effective in areas with lower rates of TB, as well as whether its use will improve clinical practice and ultimately lead to better outcomes for patients. The hope is that the test will result in more rapid diagnosis and faster treatment, reducing the number of avoidable deaths from TB meningitis. Additional Information Please access these websites via the online version of this summary at http://dx.doi.org/10.1371/journal.pmed.1001536. • This study is further discussed in a PLOS Medicine Perspective by David Boulware • The US National Institutes of Health provides information on TB meningitis • The World Health Organization provides information on tuberculosis and HIV • The US Centers for Disease Control and Prevention has a factsheet on HIV and TB • AIDS.gov also provides information about HIV and TB

The advent of HIV and AIDS has brought about many diagnostic and management challenges regarding multiple opportunistic infections. Pneumocystis jirovecii pneumonia (PCP) is a common presentation in patients with AIDS who are not on prophylaxis or highly active antiretroviral therapy (HAART). Herpes simplex 1 virus (HSV-1) is a ubiquitous virus that mainly causes benign disease during primary infection. However, it is known to cause severe pneumonia and disseminated disease in the immunocompromised.1 We present a case of HSV-1 pneumonitis and PCP co-infection in an HIV-positive patient with respiratory failure. To the best of our knowledge, based on Pubmed and Google Scholar searches, this is the first case to be reported in the English language literature.

Background The yield of mycobacterial blood cultures for multidrug-resistant (MDR) and extensively drug-resistant tuberculosis (XDR-TB) among drug-resistant TB suspects has not been described. Methods We performed a retrospective, cross-sectional analysis to determine the yield of mycobacterial blood cultures for MDR-TB and XDR-TB among patients suspected of drug-resistant TB from rural South Africa. Secondary outcomes included risk factors of Mycobacterium tuberculosis bacteremia and the additive yield of mycobacterial blood cultures compared to sputum culture. Results From 9/1/2006 to 12/31/2008, 130 patients suspected of drug-resistant TB were evaluated with mycobacterial blood culture. Each patient had a single mycobacterial blood culture with 41 (32%) positive for M. tuberculosis, of which 20 (49%) were XDR-TB and 8 (20%) were MDR-TB. One hundred fourteen (88%) patients were known to be HIV-infected. Patients on antiretroviral therapy were significantly less likely to have a positive blood culture for M. tuberculosis (p = 0.002). The diagnosis of MDR or XDR-TB was made by blood culture alone in 12 patients. Conclusions Mycobacterial blood cultures provided an additive yield for diagnosis of drug-resistant TB in patients with HIV from rural South Africa. The use of mycobacterial blood cultures should be considered in all patients suspected of drug-resistant TB in similar settings.