HIV-related tuberculosis is difficult to diagnose and is associated with high morbidity and mortality. Recently, the World Health Organization has endorsed the GeneXpert MTB/RIF (Xpert) assay for the diagnosis of pulmonary tuberculosis in HIV-infected patients from developing countries, but information about the use of Xpert for the diagnosis of extrapulmonary tuberculosis is scarce. In this study, we compared the performance of light-emitting diode (LED) auramine fluorescent microscopy and the Xpert assay for the diagnosis of tuberculosis in HIV infected patients in a district hospital of India. Although at higher cost, Xpert outperformed LED fluorescent microscopy in all type of specimens, especially in cerebrospinal fluid where the number of positive results was increased 11 times. Pleural fluid, ascitic fluid, pus, and stool specimens also yielded positive results with the Xpert assay. When collecting two additional early-morning sputum samples, the increase of the number of positive results with the Xpert assay was lower than previously reported for HIV infected patients. Rifampicin resistance was observed in 2.2% of the cases. The results of this study show that the Xpert assay can dramatically improve the rapid diagnosis of tuberculous meningitis and other types of extrapulmonary tuberculosis of HIV infected patients.

The Xpert MTB/RIF assay can detect mutations in rpoB gene that confer rifampicin resistance (RR) using five overlapping probes (A, B, C, D, and E). In this study, we described our experience with the Xpert assay in a rural setting in India. During the study period, 3250 samples were processed. The result was unsuccessful in 5.7% of cases. For extrapulmonary specimens, the risk of unsuccessful result was higher in tissue biopsy and stool samples. Among samples positive for Mycobacterium tuberculosis, rifampicin resistance was indeterminate in 1.2% of them. Our results and a review of the literature showed that the most frequent mutations conferring RR were located in the region of Probe E (63.6%; 95% confidence interval [CI] 56.26–70.94), followed by Probe B (15.02%; 95% CI 11.94–18.10), Probe D (13.35%; 95% CI 10.01–16.69), Probe A (4.73%; 95% CI 1.92–7.54), and Probe C (1.61%; 95% CI 0.67–2.54). Although the high cost of the cartridges precluded using the Xpert assay for routine diagnosis of tuberculosis, our results demonstrate that the assay can be used to diagnose RR-tuberculosis in rural areas with limited laboratory infrastructure and could be a convenient tool to investigate the molecular epidemiology of RR in resource-limited settings.

Staphylococcus aureus (SA) is the most common cause of skin and soft tissue infections (SSTIs) and nosocomial infections. In developed countries there is a major concern about the rise of community-associated methicillin-resistant SA (CA-MRSA), but data from developing countries are scarce. In this study we describe the prevalence and antibiotic susceptibility of CA-MRSA and healthcare-associated MRSA (HA-MRSA) in a district hospital from rural India. We identified 119 CA-SA infections and 82 HA-SA infections. The majority of infections were SSTI, and the proportion of MRSA in CA-SA and HA-SA infections was 64.7% and 70.7%, respectively. The proportion of CA-MRSA in children <5 years was 73.7%. We did not observe any linezolid or vancomycin resistance. CA-SA had high levels of resistance to ciprofloxacin and low levels of resistance to chloramphenicol, doxycycline, rifampicin, and clindamycin. CA-MRSA had higher proportion of resistance to ciprofloxacin, erythromycin, gentamicin, and cotrimoxazole than CA methicillin-susceptible SA (CA-MSSA). HA-MRSA had higher proportion of resistance to clindamycin and doxycycline than CA-MRSA. The results of this study indicate that MRSA is replacing MSSA in CA-SA infections. If these findings are confirmed by other studies, the spread of CA-MRSA can be a major public health problem in India.

Studies performed in developed countries have shown that infections by third generation cephalosporin resistant Escherichia coli (G3CREC) are associated with increased mortality, but data from developing countries are scarce. In this observational study, we collected clinical and microbiological information of 194 patients admitted to a district hospital in India who had community-acquired isolation of Escherichia coli. The proportion of patients with G3CREC was 79.4%. In a multivariable logistic regression analysis, factors associated with 21-day mortality were isolation from a normally sterile site, HIV infection and isolation of G3CREC. Strains of Escherichia coli isolated from normally sterile sites had lower levels of resistance to quinolones and beta-lactam antibiotics. The proportion of meropenem and ciprofloxacin resistance was 11.1% and 80.9% respectively. The high proportion of G3CREC in the community and the association of G3CREC with 21-day mortality indicate that G3CREC is a major public health problem in developing countries.

Cryptococcal meningitis (CM) is a common cause of death among HIV infected patients in developing countries, especially in sub-Saharan Africa. In this observational HIV cohort study in a resource-limited setting in India, we compared the standard two-week intravenous amphotericin B deoxycholate (AmBd) (Regimen I) with one week of intravenous AmBd along with daily therapeutic lumbar punctures and intrathecal AmB lipid emulsion (Regimen II) during the intensive phase of CM treatment. 78 patients received Regimen I and 45 patients received Regimen II. After adjustment for baseline characteristics (gender, age, altered mental status or seizures at presentation, CD4 cell count, white blood cells, cerebrospinal fluid white cells, and haemoglobin), the use of Regimen II was associated with a significant relative risk reduction in mortality (adjusted hazard ratio 0.4, 95% confidence interval, 0.22–0.76) and 26.7% absolute risk reduction (95% confidence interval, 9.9–43.5) at 12 weeks. The use of Regimen II resulted in lower costs of drugs and hospital admission days. Since the study is observational in nature, we should be cautious about our results. However, the good tolerability of intrathecal administration of AmB lipid emulsion and the clinically important mortality reduction observed with the short-course induction treatment warrant further research, ideally through a randomized clinical trial.

Enumeration of CD4 lymphocytes is essential for the clinical management of HIV-infected patients, but it can be difficult to afford in developing countries. In this study we evaluated a reagent reduction strategy for reducing the cost of enumerating CD4 cell absolute count and percentage using the FACSCalibur flow cytometer (Becton Dickinson). We compared the protocol recommended by the manufacturer with a protocol that used half of the usual amount of CD3/CD4/CD45 monoclonal antibody reagent in 100 samples from HIV-infected patients in a rural hospital in India. The concordance correlation coefficient between the two protocols was 0.976 for CD4 cell count and 0.984 for CD4 cell percentage. We did not find significant bias when performing Deming regression or Bland-Altman analysis. Sensitivity and specificity were 97% and 98.5% for identifying patients with less than 200 CD4 cells/μL, 98.1% and 93.8% for identifying patients with less than 350 CD4 cells/μL, and 100% and 94.7% for identifying patients with less than 25% CD4 cells, respectively. This reagent reduction strategy can be used for reducing the cost of enumerating CD4 lymphocytes in high-volume laboratories from resource-limited settings.

With the implementation of 2010 World Health Organization guidelines, the number of infants from developing countries who will initiate antiretroviral therapy (ART) will increase considerably. In this study we describe the HIV antibody tests of 14 HIV infected children who initiated ART at age less than one year in a rural setting of India. The HIV rapid test was negative in seven and indeterminate in two cases, whereas the HIV enzyme-linked immunosorbent assay (ELISA) antibody test was negative in three and indeterminate in one case. In one child who had both negative HIV rapid test and ELISA initially, HIV serology turned positive after having a virological failure to ART, suggesting the possibility of utilizing HIV serology for monitoring ART effectiveness in children who experience HIV seroreversion. In conclusion, HIV seroreversion of children with early initiation of ART is common and should be considered for avoiding misdiagnosis of HIV infection.

With the implementation of 2010 World Health Organization guidelines, the number of infants from developing countries who will initiate antiretroviral therapy (ART) will increase considerably. In this study we describe the HIV antibody tests of 14 HIV infected children who initiated ART at age less than one year in a rural setting of India. The HIV rapid test was negative in seven and indeterminate in two cases, whereas the HIV enzyme-linked immunosorbent assay (ELISA) antibody test was negative in three and indeterminate in one case. In one child who had both negative HIV rapid test and ELISA initially, HIV serology turned positive after having a virological failure to ART, suggesting the possibility of utilizing HIV serology for monitoring ART effectiveness in children who experience HIV seroreversion. In conclusion, HIV seroreversion of children with early initiation of ART is common and should be considered for avoiding misdiagnosis of HIV infection.