Prevalence of genotype D in chronic liver disease patients with occult HBV infection in northern region of India.

Background: Etiology of nearly 30% cases of chronic viral hepatitis remains undetected. Occult HBV infection (OBI) has emerged as an important clinical entity in this scenario. Apart from prevalence and clinical outcome of OBI patients genotype was determined in northern region of India. Materials and Methods: A total of 847 patients with chronic liver disease (CLD) were screened for common viral etiologies and others serological markers of HBV. Amplifi cation of surface, precore and polymerase genes of HBV was performed in patients negative for other etiologies. Genotyping and sequencing of the precore region was performed for OBI cases. Results: Twenty-nine (7.61%) cases of OBI were identifi edof which 9 had chronic liver disease (CHD), 11 liver cirrhosis (LC) and 9 hepatocellular carcinoma (HCC). Majority of OBI cases were detected by amplifi cation of surface gene 26 (89.6%), followed by pre-core gene 12 (41.3%). Their liver functions tests were signifi cantly deranged in comparison to overt HBV cases. IgG anti HBc was present in 8 (27.6%) OBI cases. Mutation was observed in 8 (32%) in pre-core region at nt. 1896 of overt HBV cases. Genotype D was the predominant genotype. In conclusion: OBI in our study was characterized by predominance of genotype D and more severe clinical and biochemical profi le in comparison to overt HBV. IgG anti HBc positivity could be utilized as a marker of OBI. We recommend use of sensitive nested PCR for diagnosis of OBI, amplifying at least surface and precore gene.

Comparison of the activities of amphotericin B, itraconazole, and voriconazole against clinical and environmental isolates of Aspergillus species.

Background: Invasive fungal infections are a significant cause of morbidity and mortality in immunocompromised populations. Aims: To evaluate the susceptibility pattern of our isolates against amphotericin B, itraconazole, and voriconazole and to compare the antifungal activities of these agents with each other against the Aspergillus species tested. Settings and Design: A prospective study was designed to include clinical and environmental isolates of Aspergillus species. Materials and Methods: 420 sputum samples, 70 bronchoalveolar lavage fluids, 160 oral washings, and 47 environmental samples were collected. Direct microscopy by potassium hydroxide and lactophenol cotton blue mounts followed by culture on Sabourad`s dextrose agar (SDA) was done. Susceptibility testing was performed by the broth microdilution technique as per Clinical Laboratory Standards Institute standards (M-38A). Additionally, all the isolates were also tested by the colorimetric microdilution technique using Alamar Blue dye. Statistical Analysis: It was done by the Chi-square test and Z-test using SPSS statistical software version 12.0. Results and Conclusion: Twenty-seven isolates (47.3%) were recovered from patients with chronic bronchial asthma followed by fibrocavitary pulmonary tuberculosis in 9 (15.7%), allergic bronchopulmonary aspergillosis (ABPA) in 6 cases (10.5%), bronchiectasis in 3 (5.2%), bronchogenic carcinoma in 5 (8.7%) and those receiving radiotherapy for head and neck cancer 7 (12.2%). Thirteen environmental isolates were also included in the study. The most common isolate was A. fumigatus 28 (40%), followed by A. niger 22 (31%), A. flavus 13 (19%), and A. terreus 7(10%). All isolates were susceptible to amphotericin B, itraconazole, and voriconazole. Among the three agents tested, voriconazole exhibited lowest MICs (≤1 μg/ml) against all Aspergillus species.

Acute lower respiratory tract infections in children.

Acute lower respiratory tract infection (ALRTI) is a common illness, but there have been relatively few studies of the bacterial etiology in developing countries. Nasopharyngeal aspirates of 70 children under 10 years of age with ALRTI were cultured for aerobic bacterial pathogens. Klebsiella pneumoniae was the commonest organism (32.2%) isolated followed by S. pneumoniae (10%), E. coli (10%), P. aeruginosa (5.7%), S. aureus (2.8%) and H. influenzae (1.4%). There were significantly more bacterial pathogens isolated in children <1 year of age (73.7%) than in those >1 year of age (56.2%) (P=0.03). A shift in spectrum from Gram-positive cocci to Gram-negative bacilli in ALRTI was observed in our study.

Evaluationof ParaSight F Test in Dignosis of Plasmodium Falciparum Infection.

P. falciparum malaria is a severe form of disease which requires urgent diagnosis and treatment to save the life of patient. Blood smear examination is the commonest method used for diagnosis. The present study was done to evaluate ParaSight F test in patients of P. falciparum infection.The study was performed on 100 patients who where clinically diagnosed as cases of P. falciparum infection. ParaSight F test and Leishman stained blood smear examination was done in all 100 patients (50 patients of cerebral malaria + 50 patients of acute malaria).ParaSight F test was positive in 45 patients and blood smear positive in 28 patients of cerebral malaria. 35 patients of acute malaria were positive by ParaSight F test while blood smear was positive in 15 patients. Sensitivity, specificity, positive and negative predictive values of ParaSight F test are 95.7%, 100%, 100%, 100%, 60% in cerebral malaria and 100% each in acute malaria. ParaSight F test can be used as diagnostic tool in cases of P. falciparum infections, where blood smear is negative.

Comparison of different diagnostic techniques in Plasmodium falciparum cerebral malaria.

BACKGROUND & OBJECTIVES: Plasmodium falciparum cerebral malaria remains a major health problem in India. The efficacy of treatment of cerebral malaria lies in its early diagnosis through rapid diagnostic methods. ParaSights-F test detects HRP-2 antigen secreted by parasitised red blood cells and quantitative buffy coat assay (QBC) is examination of buffy coat for the presence of malarial parasite stained with acridine orange. This study was performed to evaluate the effectiveness of ParaSight-F test and QBC assay as diagnostic methods in the patients of cerebral malaria. METHODS: Fifty clinically diagnosed patients of cerebral malaria were included in the study. ParaSight-F test, QBC and conventional blood smear examination was done. Patients who were in coma and there were no obvious features of bacterial or viral etiology were investigated for cerebral malaria by these diagnostic methods. RESULTS: ParaSight-F test, QBC and peripheral blood smears were examined. Patients were followed-up for signs of clinical recovery. ParaSight-F test was positive in 47 patients, QBC in 46 while blood smear examination was positive in 28 cases. INTERPRETATION & CONCLUSION: Sensitivity and specificity of ParaSight-F test were found to be 96.6 and 94% while QBC showed 97.8 and 100% respectively. ParaSight-F test and QBC were found to be novel methods for diagnosis of cerebral malaria especially in the cases where diagnosis can not be made by conventional blood smear examination due to low parasitaemia. These rapid diagnostic methods help in early therapeutic intervention.

Chlamydia trachomatis infection & female infertility.

BACKGROUND & OBJECTIVES: Chlamydia trachomatis is a well recognized sexually transmitted pathogen. Besides its potential to produce genital tract infection, C. trachomatis is increasingly being associated with long-term complications like infertility. The present study was undertaken to assess the role of C. trachomatis in female infertility as such data are lacking. METHODS: Women of primary and secondary infertility (n=110) and 30 healthy term pregnant women as control group were enrolled in the study. Detailed clinical history of each patient was recorded. Hysterosalpingography was performed in all patients. Endocervical swabs were collected for culture on cycloheximide treated McCoy cell line and for antigen detection by ELISA. RESULTS: C. trachomatis was detected in 31 (28.1%) of the 110 infertile women while one (3.3%) in control group was positive for C. trachomatis (P<0.01). Cell culture alone identified 25 (22.72%) patients suffering from chlamydial infection while C. trachomatis antigen was detected by ELISA in 18 (16.37%) patients. The one control case was positive for Chlamydia antigen by ELISA and not by cell culture. Chlamydial positivity was seen in 20 of the 74 (27%) women with primary infertility and in 11 of the 36 (30.6%) with secondary infertility. Of the 58 asymptomatic women, 21(36.2%) had chlamydia infection while among the 52 symptomatic cases 10 (19.2%) were infected; 38 per cent women with chlamydial infection also had tubal occlusion. INTERPRETATION & CONCLUSION: A significantly high rate of C. trachomatis infection was found in infertile women and more so in asymptomatic females and in secondary infertility cases. Lack of symptoms make clinical diagnosis of chlamydial infection difficult. Screening of infertile women for C. trachomatis is therefore recommended so far early therapeutic interventions.

Efficacy of tyndalized Lactobacillus acidophilus in acute diarrhea.

OBJECTIVE: A double-blind randomized controlled-trial was done to evaluate the efficacy of tyndalized Lactobacillus acidophilus in acute diarrhea. METHODS: All children from 6 months to 12 years with acute diarrhea were included. Lactobacillus acidophilus/placebo was given to the children for 3 days with ORS and feeds. Intake-output was recorded 4 hourly. Of the 98 children, 48 received lactobacillus and 50 the placebo. RESULTS: ORS consumed, frequency of stools, duration of diarrhea, time for rehydration, hospital stay, weight gain and IVF needed were comparable in the two study groups. There were 4 treatment failures in the lactobacillus group and none in the placebo group (OR 0.92, 95%CI 0.84-0.99). In the rotaviral diarrhea and in those who had diarrhea of less then 60 hours the difference did not reach statistical significance. CONCLUSION: There is no significant benefit of tyndalized Lactobacillus acidophilus in acute diarrhea.

Resistance due to aminoglycoside modifying enzymes in Pseudomonas aeruginosa isolates from burns patients.

BACKGROUND & OBJECTIVE: Enzymatic modifications of aminoglycosides result in high-level resistance in numerous bacterial species. However, the data on this aspect are elementary in our country. The present study was therefore designed to determine resistance rates and patterns, and to find out the prevalent aminoglycoside modifying enzymes (AMEs) in clinical isolates of Pseudomonas aeruginosa from hospitalized burn patients. METHODS: Forty two, non-repeat, clinical isolates of P. aeruginosa obtained during a period from February to July 2003, were analysed for the presence of antibiotics resistance. On the basis of aminoglycoside susceptibility patterns, resistance phenotypes and possible AMEs were inferred according to interpretative reading. Seven isolates collected during the same period and previously characterized to harbour a 48.5 kb plasmid, encoding multiple drug resistance, were also analysed for aminoglycoside susceptibility patterns, and AMEs encoded by the plasmid were inferred. RESULTS: Ninety six per cent of the isolates were multi drug-resistant and majority (71.4%) were resistant to 5 or more antibiotics. Markedly high resistance to tobramycin (83.6%) and amikacin (55.1%) was noted, whereas gentamicin resistance was present in 32.6 per cent isolates. The enzyme N-acetyl transferases (AAC) viz. AAC(6')-I was the most common isolated AME followed by AAC(3)-II in 42.8 and 20.4 per cent of isolates respectively. The plasmid harbouring isolates belonged to AAC(6') phenotype and the enzyme encoded was inferred to be AAC(6')-I. INTERPRETATION & CONCLUSION: Markedly high resistance to tobramycin and amikacin was noted in the present study. AAC(6')-I was the most common AME and was inferred to be plasmid encoded in R-plasmid harbouring isolates. This is among the premier reports regarding the aminoglycoside resistance due to AMEs especially plasmid encoded, in P. aeruginosa from India. Further studies are required from different parts of the country to findout the prevalence of aminoglycoside resistance due to AMEs in P. aeroginosa isolates.

Age-specific prevalence of hepatitis B surface antigen in pediatric population of Aligarh, North India.

OBJECTIVE: To estimate the age-specific seroprevalence of hepatitis B surface antigen (HBsAg) in children upto 14 years of age. METHODS: Equal number (115 each) of apparently healthy children of both sexes of different age groups i.e. < 1, 1-4, 5-9 and 10-14 years, attending pediatric outpatient services and Well Baby Clinic of the hospital were tested for HBsAg using ELISA test. Positive results were confirmed by a second ELISA. RESULTS: Overall 4.35% (95%CI, 2.44 - 6.25) of the 460 children tested were HBsAg positive. The prevalence rate was the highest (6.09%) in the 1-4 year age category. In the < 1, 5-9 and 10-14 year age groups it was 4.35%, 4.35% and 2.61% respectively. The overall male to female ratio was 2.1:1, with no significant difference in seropositivity rates (P = 0.816).The difference in the prevalence rates between the rural (4.84%) and urban populations (3.77%) was also statistically insignificant (P = 0.577). CONCLUSION: Average HBsAg positivity in the pediatric population in this region is 4.35% (95%CI, 2.44 - 6.25). The prevalence progressively increases and peaks in the 1- 4 years age group. It is least in 10-14 years age group.

Antibody levels against polioviruses in children following Pulse Polio Immunization Program.

This cross sectional study was performed in a tertiary level teaching hospital to evaluate and compare the antibody levels in children below 6 years who had received oral polio vaccination through Pulse Polio Immunization (PPI) with those children who had received both routine immunization as well as PPI. Detail history of polio immunization was taken. Serum samples were then collected for antibody determination by neutralization tests with standard polio viruses using Vero cell lines. Total 400 children were studied; 14 were found unvaccinated. Out of the remaining 386 (96.5%) vaccinated children, 292 (75%) had received both routine and pulse polio immunization, 68 (17%) had only PPI while 26 (6.7%) had received only routine immunization. The seropositivity was lowest for P3 and highest for P2. Overall seroprevalence for PI, P2 and P3 in vaccinated children was 89.1%, 93% and 80.6% respectively, and did not differ significantly between the three vaccinated subgroups. However, children who were immunized by both routine and PPI had higher geometric mean titers (315.5, 484.7 and 187.4 for PI, P2 and P3 respectively) when compared with those who had received only PPI (P<0.001 for each PI, P2 and P3), as well as those who had received only routine immunization with OPV (P<0.05 for PI, p<0.001 for P2, and P<0.01 for P3). Despite the reasonable immunization coverage in study population, there were 29 (7.25%) triple negative cases. Hence other causes of low seroconversion should also be considered to achieve polio free India.

Prevalence of human immunodeficiency virus infection in children with tuberculosis.

This prospective study was carried out in the pediatric ward and outpatient department of a tertiary care centre to estimate the prevalence of HIV seropositivity in children with tuberculosis. Two hundred and fifty consecutive children below 12 years of age with (pulmonary and extrapulmonary) tuberculosis diagnosed between March 1999 and July 2000 were screened for HIV infection. A patient was labeled as HIV positive if two consecutive ELISA tests were found positive using different antigen/principle. Supplemental western blot test was also done. Parents of seropositive children were also screened for HIV infection and tuberculosis. Total 5 cases were HIV positive giving a seroprevalence of 2%. All the five patients had disseminated tuberculosis. We suggest regular screening of children with disseminated/miliary tuberculosis for HIV co-infection.

Prevalence of aspergillosis in bronchogenic carcinoma.

Bronchoalveolar lavage of 42 patients of bronchogenic carcinoma was studied to find out the prevalence of aspergillosis. Sera of the patients were also analysed for presence of anti-Aspergillus antibodies by Immunodiffusion (ID), Enzyme linked immunosorbent assay (ELISA) and dot blot assay (DBA). Aspergillus was isolated in culture from 6 (14.2%) patients of bronchogenic carcinoma. Aspergillus fumigatus was the predominant species isolated. All the strains of Aspergillus were sensitive to itraconazole, ketoconazole and amphotericin B while resistance (33.3%) was found with fluconazole. Anti-aspergillus antibodies were detected equally by ID, ELISA and DBA in 9 (21.4%) cases. The present study revealed prevalence and seroprevalance of Aspergillus in bronchogenic carcinoma to be 14.2% and 21.4% respectively. Consistent reactivity against 18 kDa Aspergillus fumigatus antigen was noted in serologically positive cases. Antibodies against 18 kDa protein antigen in western blotting may be used as a reference marker for diagnosis of aspergillosis in bronchogenic carcinoma. It is also suggested that the simplest serological technique like ID may be performed along with culture for diagnosing Aspergillosis in patients of bronchogenic carcinoma since ID, ELISA and DBA showed similar sensitivity.