Antiretroviral treatment, viral load of mothers & perinatal HIV transmission in Mumbai, India.

Background & objectives: Mother-to-child transmission (MTCT) is the most significant route of HIV transmission in children below the age of 15 yr. In India, perinatal HIV transmission, even after treatment, accounts for 5.4 per cent of HIV cases. The present study was conducted to evaluate the efficacy of anti-retro viral therapy (ART) or prophylactic treatment (PT) to control maternal viral load in HIV positive women, and its effect on vertical HIV transmission to their infants. Methods: A total of 58 HIV positive women were enrolled at the time of delivery and their plasma samples were obtained within 24 h of delivery for estimation of viral load. Viral load analysis was completed in 38 women. Infants received single dose nevirapine within 2 h of birth and zidovudine for 6 wk. At the end of 18 month follow up, HIV positive or negative status was available in 28 infants. Results: Results revealed undetectable levels of viral load in 58.3 per cent of women with ART compared to 30.7 per cent of women with PT. No women on ART had viral load more than 10,000 copies/ml, whereas seven (26.9%, P=0.07) women receiving PT had this viral load. Median CD4 count of women on PT (483 cells/μl) was high compared to the women on ART (289 cells/ μl). At the end of 18 months follow up, only two children were HIV positive, whose mothers were on PT. One had in utero transmission; infection detected within 48 h of delivery, while the other child was infected post partum as HIV was detected at six months follow up. Interpretation & conclusions: Women who received a single dose of nevirapine during delivery had higher levels of viral load than women on ART. Combination drug therapy for pregnant women is now a standard of care in most of the western countries; use of nevirapine monotherapy at the time of delivery in our settings is not effective in controlling viral load. This highlights initiation of ART in pregnant women to control their viral load and thus to inhibit mother to child HIV transmission.

Polymorphisms in major cytokine genes: A study among human immunodeficiency Virus-1 serodiscordant couples in Mumbai, India.

Purpose: Polymorphism in cytokine genes may affect its production, which play an important role in modulation of human immunodeficiency virus (HIV) infection. Evaluation of these polymorphisms might help to understand why some individuals remain uninfected in spite of several exposures to HIV infection, such as the negative spouses of discordant couples. The aim of this study was to evaluate the association of 22 single nucleotide polymorphisms (SNPs) in 13 cytokine genes and their receptors with HIV infection in serodiscordant couples, attending the Integrated Counselling and Testing Centre of a Municipality Hospital. Materials and Methods: At the end of at least 2 years of follow up, 42 couples were confirmed as being serodiscordant. Genotyping was carried out in blood samples of these couples using the polymerase chain reaction-sequence-specific amplification method. Results: Significantly high frequency of interleukin-1 receptor antagonist IL-1RA mspa 11100 CC ( P = 0.04), tumor necrosis factor-alpha TNF-α −238 AG ( P = 0.01) and IL-4 −33 TT ( P = 0.01) was observed in HIV seropositives (HSP) while frequency of TNF-α −238 GG ( P = 0.02) was significantly high among the exposed uninfected (EU). However, application of Bonferroni correction identified only two SNPs i.e., TNF-α −238 AG and IL-4 −33 TT to be significantly associated with the acquisition of HIV. In remaining cytokine genes, no significant association was observed. Conclusion: Our study highlighted possible association of certain specific polymorphisms with HIV transmission, whereas presence or absence of certain other polymorphism in EU individuals might be offering protection from HIV infection. These variations at the genetic level might help to explore new insights into treatment and HIV prevention strategies.

Evaluation of Calypte AWARE HIV-1/2 OMT antibody test as a screening test in an Indian setting.

Purpose: Integrated counselling and testing centres (ICTC) provide counselling and blood testing facilities for HIV diagnosis. Oral fluid tests provide an alternative for people whodo not want blood to be drawn. Also, it avoids the risk of occupational exposure. The goal of this study was to evaluate the utility of Calypte AWARE HIV-1/2 OMT antibody test as a screening test in an Indian setting. Materials and Methods: A cross-sectional study was carried out after ethics committee approval in 250 adult ICTC clients. Blood was collected and tested from these clients for HIV diagnosis as per routine policy and the results were considered as the gold standard. Also, after another written informed consent, oral fluid was collected from the clients and tested for the presence of HIV antibodies. Twenty five clients who had and 25 clients who had not completed their secondary school education (Group A and Group B, respectively) were also asked to perform and interpret the test on their own and their findings and experiences were noted. Result: The sensitivity, specificity, PPV and NPV of the oral fluid antibody test were 100%, 98.51%, 94.11% and 100%, respectively. Seventy six percent of clients preferred oral fluid testing. Group B found it difficult to perform the test as compared to Group A and this difference was statistically significant (P ≤ 0.05). Conclusion: Oral fluid testing can be used as a screening test for HIV diagnosis; however, confirmation of reactive results by blood-based tests is a must.

Study of HIV-1 subtypes in serodiscordant couples attending an integrated counselling and testing centre in Mumbai using heteroduplex mobility analysis and DNA sequencing.

Aims: To determine the prevalent subtypes of HIV-1 in serodiscordant couples. Setting: Integrated Counselling and Testing Centre (ICTC), Department of Microbiology. Study Design: Prospective pilot study. Participants: Thirty HIV-1 serodiscordant couples. Inclusion Criteria: a) Documentation of HIV-1 infection in one partner and seronegative status in the other, current history of continued unprotected sexual activity within the partnership, demonstration that they have been in a partnership for at least 1 year and are not currently on highly active antiretroviral therapy HAART; b) willingness of both partners to provide written informed consent including consent to continued couple counselling for 3 months. Materials and Methods: HIV-1 subtyping was carried out by heteroduplex mobility analysis (HMA) by amplifying env region; and DNA sequencing by amplifying gag region. Results: HIV-1 env gene was amplified successfully in 10/30 samples; gag gene, in 25/30 samples; and both env and gag gene were amplified successfully in 5/30 samples. HIV-1 subtype C was detected from 21 samples; subtype B, from 7; and subtype A, from 2. Sample from 1 positive partner was detected as subtype C by env HMA and subtype B by gag sequencing. Conclusion: HIV-1 subtype C was found to be the predominant subtype of HIV-1 in serodiscordant couples attending our ICTC, followed by HIV-1 subtype B and HIV-1 subtype A, respectively. DNA sequencing was found to be the most reliable method for determining the subtypes of HIV-1.

Enterococcal infections with special reference to phenotypic characterization & drug resistance.

BACKGROUND & OBJECTIVES: Enterococci, classified as group D streptococci, are the second leading cause of nosocomial infections. The incidence of enterococcal infections and species prevalent in India is not thoroughly investigated. The present study was undertaken to isolate and characterize enterococci from clinical specimens and determine the antimicrobial susceptibility pattern of these isolates. METHODS: Clinical specimens (blood, urine and swabs) were cultured on bile esculin azide agar (BEAA) for isolation of enterococci. The phenotype based scheme included Gram staining of growth on BEAA and subculturing of cocci on sheep blood agar plates for vancomycin disk diffusion and hydrolysis of L-pyrrolidonyl-beta-napthylamide (PYR) testing. The phenotypic method was used to surveillance cultures that yielded growth on BEAA. Enterococcal strains were further identified to the species level by conventional biochemical tests. PYR positive isolates were further characterized into vancomycin resistant enterococci (VRE) and nonVRE depending upon vancomycin inhibition zone size. The isolates were characterized into vanA, vanB and vanC depending upon minimum inhibitory concentration (MIC) values. Conventional method was used to study the antibiogram of isolates. RESULTS: A total of 52 isolates of enterococci (10 Enterococcus faecalis, 42 E. faecium) were isolated from 534 clinical specimens. Of the 52 isolates, 12 isolates were resistant to vancomycin with an MIC > 4 microg/ml but sensitive to teicoplanin (vanB isolates). INTERPRETATION & CONCLUSION: Our study reveals the problem of multiple drug resistant enterococci and emergence of VRE. Better susceptibility tests need to be used to measure the vancomycin resistance accurately.

Persistent strongyloidiasis in a case of common variable immunodeficiency.

A case of strongyloidiasis in common variable immunodeficiency is reported here. The patient was given several courses of albendazole without any response.

Seroprevalence of IgG antibodies to Helicobacter pylori among gastrointestinal endoscopists, radiologists and paramedical personnel.

BACKGROUND: The route of transmission of Helicobacter pylori is unknown. Since the organism has been isolated from saliva, gastric juice and stool, medical personnel could be at high risk for acquiring the infection during procedures like gastrointestinal endoscopy. AIMS: To study whether endoscopy is a professional hazard for acquisition of H. pylori. METHODS: We studied the prevalence of IgG antibodies to H. pylori in endoscopists (n = 17), radiologists (n = 17) and personnel from paraclinical branches (n = 35); microbiology (n = 21), pathology (n = 7) and forensic medicine (n = 7); among the paraclinical personnel five were at high risk because they worked with cultures of H. pylori. Subjects answered a questionnaire regarding upper gastrointestinal symptoms, and precautions taken at the work place against infection. The serum was tested for IgG antibodies to H. pylori using a microwell ELISA and a rapid card test. RESULTS: H. pylori antibodies were present in five (29.4%) endoscopists, three (17.6%) radiologists and seven (20%) paraclinical personnel; only one of the 5 high risk para medical personnel was positive. There was no correlation between the duration of performing endoscopies and the H. pylori IgG status. CONCLUSION: Endoscopy is not a risk factor for acquiring H. pylori infection.

Dental plaque: an unlikely reservoir of Helicobacter pylori.

BACKGROUND AND AIM: Dental plaque has been suggested as a reservoir for Helicobacter pylori, though data in this regard are conflicting. We evaluated the prevalence of H. pylori DNA in dental plaque using polymerase chain reaction (PCR). METHODS: Antral H. pylori status of 156 patients with acid-peptic disease (APD) was studied by rapid urease test (RUT), histology and culture. Dental plaque obtained from these 156 patients and 92 healthy volunteers was evaluated for the presence of H. pylori using RUT, culture and PCR. RESULTS: H. pylori was present in 133 antral biopsy samples by RUT and/or histology. The dental plaque of 37 patients with APD and 21 healthy volunteers tested positive by RUT. H. pylori was not isolated by culture from any of the dental plaques. PCR gave a significant amplification product in 11 of 248 (4.4%) dental plaque samples, 7 from patients with APD and 4 from normal healthy volunteers. CONCLUSION: The frequency of H. pylori in the dental plaque is low, and this is unlikely to be a prominent site of infection with H. pylori.

Epidemiology of Helicobacter pylori: the Indian scenario.

OBJECTIVE: To compare the age-related prevalence of Helicobacter pylori in populations from developing and developed nations to determine the possible mode of transmission. METHODS: Endoscopic gastric biopsies (for biopsy urease test and histology) were obtained in 526 patients with dyspepsia and in 82 control subjects to determine H pylori prevalence. RESULTS: H pylori prevalence in patients with dyspepsia and in control subjects was 65% and 46% respectively. Age-related prevalence in these two groups in the age groups 10-19 years, 20-29 years, 30-39 years, 40-49 years and > or = 50 years was 52%, 70%, 69%, 60% and 59%, and 44%, 55% 58%, 36% and 33% respectively. CONCLUSION: Exposure to H pylori occurs early in India and is widespread, even in control subjects. The high prevalence of the organism in young Indian control subjects and the comparable prevalence of antibodies to H pylori and hepatitis A virus infection in different age groups both in developed and developing nations may suggest a feco-oral mode of transmission.

Mono and dual therapy for Helicobacter pylori associated gastritis.

Sixty patients with Helicobacter Pylori positive non ulcer dyspepsia were randomly allocated to one of the following treatment groups: Group I--norfloxacin 400 mg bid for 10 days, Group II--amoxycillin 500 mg bid plus tinidazole 500 mg bid for 15 days, Group III--colloidal bismuth subcitrate (CBS) 240 mg bid for 4 weeks. H pylori elimination was achieved in 14%, 81%, and 62% in Groups I, II and III respectively. Eradication of H pylori was not observed in Groups I and II, but was achieved in 25% of patients in Group III. Antral gastritis improved in 69% in Group II and 50% in Group III. We conclude that norfloxacin is not effective in H pylori infection. A combination of amoxycillin and tinidazole is highly effective in H pylori elimination with improvement in associated gastritis, but H pylori eradication is not observed with this therapy. CBS is also effective in H pylori elimination though H pylori eradication is achieved in only 25%.