Detection of respiratory syncytial virus & Mycoplasma pneumoniae in paediatric lower respiratory tract infections

Background & objectives: Respiratory syncytial virus (RSV) and Mycoplasma pneumoniae are considered common cause of lower respiratory tract infections (LRTIs) in children. The present study was conducted to detect M. pneumoniae and RSV in paediatric LRTIs employing serology, polymerase chain reaction (PCR) and reverse transcriptase PCR (RT-PCR) analysis. Methods: Seventy five children aged one month to five years with acute LRTIs were investigated for M. pneumoniae antibodies and RSV antigen using immunochromatographic test, RT-PCR for RSV and M. pneumoniae by PCR on nasopharyngeal aspirates. Results: RSV infection was observed in 33 (44%) and M. pneumoniae was positive in 26 (35%) children. No significant difference in infection was noted between male and female children. Clinical and radiological features among RSV and M. pneumoniae positive and negative cases were similar. Considering RT-PCR for RSV as gold standard, RSV antigen immunochromatography was 90.90 per cent sensitive and 100 per cent specific. Interpretation & conclusions: Our study showed the presence of RSV and M. pneumoniae infection in 44 and 35 per cent children, respectively with community-acquired LRTIs and aged less than five years.

HLA-DQA1 & DQB1 variants associated with hepatitis B virus-related chronic hepatitis, cirrhosis & hepatocellular carcinoma

Background & objectives: Clinical outcome after hepatitis B virus (HBV) exposure varies extremely from spontaneous clearance to chronic hepatitis B and often progresses to liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Host genetic factor plays an important role in the regulation of immune response. This study was aimed to investigate whether HLA class II DQA1 and DQB1 gene polymorphism were associated with chronic hepatitis B infection and in the development of HBV-related LC and HCC. Methods: DQA1 and DQB1 allele polymorphism were studied in 187 patients with HBV-related liver diseases (which included 73 chronic hepatitis B, 84 LC and 30 HCC patients) and 109 controls who had spontaneously recovered from HBV infection using polymerase chain reaction amplification with sequence-specific primers. Results: Our data suggested that DQA1*0101/2/4 [odds ratio (OR)=2.78; Pc=0.003], DQA1*0103 (OR=2.64; Pc=0.0007) and DQB1*0302/3 (OR=2.15; Pc=0.01) were associated with the protection from chronic HBV infection, whereas DQB1*0402 (OR=0.25; Pc=0.001) showed susceptible effect on chronic HBV infection. DQB1*0601 (OR=3.73; Pc=0.006) conferred protective effect from developing LC; similarly, DQB1*0302/3 (OR=5.53; Pc=0.05) and DQB1*0402 (OR=0.00; Pc=0.001) conferred protective effect from developing HCC. However, DQA1*0601 and DQB1*0503 showed susceptible effect on chronic HBV infection; these associations were no longer significant after Bonferroni correction. Interpretation & conclusions: Our results revealed HLA-DQA1*0101/2/4 - DQA1*0103 - DQB1*0302/3 and DQB1*0601 as protective and DQB1*0402 as risk alleles. The study suggests that various subtypes of HLA-DQA1 and DQB1 are associated with both HBV clearance and development of chronic HBV infections.

A study on gender‑related differences in laboratory characteristics of dengue fever.

Studies have reported significant gender‑related differences in serological tests for detection of NS1 antigen and IgM antibody used for diagnosing dengue fever. However, no such study has been undertaken in India though dengue fever is endemic in this country. Therefore, this study was planned to study the association of serological findings with gender in 700 patients suspected to be suffering from dengue fever in the Indian setting. Haematological parameters of seropositive patients were also studied. Seropositivity and haemorrhagic findings were significantly associated with the female gender. Positive NS1 antigen and IgM antibody results were significantly associated with females and males, respectively.

Rising trend of antimicrobial resistance among Neisseria gonorrhoeae isolates and the emergence of N. gonorrhoeae isolate with decreased susceptibility to ceftriaxone.

Context: Gonorrhoea is one of the most common sexually transmitted infections (STI) in developing countries and is a global health problem. Aims: To analyze the trend of antimicrobial susceptibility of Neisseria gonorrhoeae isolates over the years, in a tertiary care hospital of North India. Settings and Design: The study population comprised males with urethritis and females with endocervicitis attending STI clinic of our hospital. Materials and Methods: In our STI laboratory, all gonococcal isolates are subjected to antimicrobial susceptibility testing by disc diffusion method as per CLSI guidelines. ß-lactamase production is determined by chromogenic cephalosporin test. Minimum Inhibitory Concentration (MIC) for ceftriaxone is determined by E-test. Statistical Analysis Used: Data were expressed as percentages. The differences in percentages were tested for statistical signifi cance by using χ2 test and P values were determined. Results: The percentage of penicillinase producing N. gonorrhoeae (PPNG) increased from 8% in 1995-96 to 20% in 2004-05 and 88% in 2011-2013. Quinolone-resistant N. gonorrhoeae (QRNG) showed a signifi cant increase from 12% in 1995-96 to 98.3% in 2004-05, while 84% isolates were found to be QRNG by 2011-2013. In January 2013 we detected our fi rst gonococcal isolate with decreased susceptibility to third-generation cephalosporins; Ceftriaxone, Cefi xime and Cefpodoxime (MIC for ceftriaxone = 0.19 μg/ml). Conclusions: The results of our study highlighted an alarming increase in the percentage of PPNG and QRNG strains over the years. Emergence of N. gonorrhoeae isolates with decreased susceptibility to third-generation cephalosporins is a cause of concern and thus emphasises the importance of antimicrobial susceptibility testing.

A study of changing trends of prevalence and genotypic distribution of hepatitis C virus among high risk groups in North India.

Purpose: Hepatitis C virus (HCV) has emerged as a leading cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma. There is a great variability in HCV’s geographical presence, transmission routes, genotypic distribution etc., in studied populations. We undertook this study in a North Indian hospital on patients of chronic liver disease to observe any emerging trend in risk groups, transmission patterns, genotypic distribution of the virus in this geographical region and its correlation with viral load. Materials and Methods: There were 54 anti-HCV positive patients including 31 HCV Ribonucleic acid (RNA) positive patients were included in the study. HCV genotyping was carried out by restriction fragment length polymorphism (RFLP) followed by direct sequencing of the core region. Viral load estimation was carried out by Taqman real time polymerase chain reaction system. Results: In 45/54 (83.3%) anti-HCV positive patients, iatrogenic procedures were responsible for transmission with blood transfusion alone responsible in 36/54 (67%). Genotype 3 was observed to be the commonest type found in all risk groups followed by type 1 and 2. Subtype 3b (35.5%) was found more prevalent than subtype 3a. A higher frequency of subtype 1b (19.4%) was also seen. Genotype 1 was associated with a signifi cantly higher viral load compared to genotypes 3 and 2. No signifi cant difference was observed in the biochemical profi le among the three genotypes except for the levels of the enzyme, aspartate aminotransferase (AST). Conclusions: Iatrogenic procedures, especially contaminated blood transfusion etc., still contributes signifi cantly to the pool of HCV infection. Genotype 3 is the predominant genotype in North India, though the subtype distribution within genotype 3 may be changing. The association of severe liver disease is signifi cantly more with genotype 1 as evidenced by higher viral load and deranged AST levels.

Awareness of changing trends in epidemiology of dengue fever is essential for epidemiological surveillance.

Dengue has become endemic in India with outbreaks occurring almost every year. The seroprevalence and serotypic data of the last 7 years in samples obtained from suspected dengue patients from a tertiary care hospital were analyzed. Out of 7846 serum samples received in the virology laboratory from suspected dengue cases during 2002 to 2008, 2366 (30.15%) were serologically confirmed. Serotyping was done using mRT-PCR. All the four serotypes were detected in 2003, while data in 2004, 2005 and 2006 revealed the the predominance of Den-3. In the year 2007 predominance of Den-2 was observed, whereas in 2008 Den-1 was the most common serotype isolated. Overall, Den-2 and Den-3 were the most predominant serotypes during 2003-2007 but Den-1 replaced these strains in the year 2008. Since the emergence of a new predominant strain can lead to the occurrence of an outbreak, presence of Den-1 in the year 2008 would pose an alarming situation before us. Well-targeted population-based epidemiological studies are urgently required to control dengue menace.

Cytomegalovirus infection: an Indian perspective.

Human cytomegalovirus (CMV) poses an important public health problem as it may cause serious morbidity and mortality in congenitally infected newborns and immunocompromised patients, most notably transplant recipients and HIV-infected persons. It is probably one of the most common infections known to humans and is characterized by a self-limiting infection in healthy individuals. CMV infection is the single most frequent cause of infectious complications in the early period following kidney transplantation Post-transfusion cytomegalovirus infection is of concern in the immunocompetent as well as in certain categories of immunocompromised individuals such as neonates, pregnant women, recipients of bone marrow and other organ transplants and individuals with immunodeficiency disorders. The emergence of AIDS in India has necessitated the establishment of reliable tests for diagnosis of cytomegalovirus infection as a damaged immune system permits cytomegalovirus reactivation. The magnitude of this problem in India and the various diagnostic modalities used have not been adequately investigated and, hence, CMV infection is still a major health problem warranting strong preventive measures. The ultimate goal of the prevention program is to develop a vaccine that can be administered to seronegative women of childbearing age to prevent primary infection during pregnancy.

Primary tuberculosis of tonsil and posterior oropharyngeal wall.

Pharyngeal tuberculosis is rare and usually occurs in association with primary pulmonary disease. Primary tuberculosis involving the palatine tonsils and the posterior oropharyngeal wall is still a rare clinical entity. We report one such case of primary tuberculosis involving both the palatine tonsils and the posterior oropharyngeal wall in a 22 year-old male. The patient responded to anti-tubercular treatment with complete disappearance of lesion and no sign of recurrence on one year follow-up. The final diagnosis was based upon histopathological report.

Lupus Vulgaris of external nose.

Lupus vulgaris is the most common morphological variant of cutaneous tuberculosis. In India, the most common sites of involvement are trunk and buttocks. The involvement of other sites like nose, periocular region and face, have been occasionally reported from India. This report presents an unusual case of lupus vulgaris involving the external nose. The diagnosis was confirmed on histopathology of the deep biopsy taken from an ulcerated part of the lesion. The response to six months of anti- tubercular treatment was dramatic.

Hepatitis B vaccine in the EPI schedule.

OBJECTIVE: This study was carried out to evaluate the adequacy of seroconversion when Hepatitis B vaccine is given along with other vaccines at 0, 6 weeks (along with DPT and OPV) and at 9 months (along with measles). METHODS: 725 infants born to apparently healthy mothers were enrolled to receive the hepatitis B vaccine at 0, 6 weeks and 9 months (Group A) or at 0, 1 and 6 months as per WHO schedule--(Group B). Baseline HbsAg testing was carried out and the babies were immunized with the first dose of hepatitis B vaccine within 48 hours of birth. BCG and the other EPI vaccines were given as per schedule. Serum samples were collected 4 weeks after the second and the third immunizations. 604 infants (83.3%) completed the study. The testing for HbsAg and Anti Hbs titers were conducted in the Department of Microbiology, Maulana Azad Medical College, New Delhi utilizing standard ELISA kits. RESULTS: The seroconversion rates 4 weeks after the second dose of the vaccine were 90.89% (GMT = 48.23) and 91.82% (GMT = 43.95) (P=0.8) in Group A and Group B respectively. After 4 weeks of the third dose the seroconversion rates were 98.99 (GMT = 161.12) and 98.45 (GMT = 150.12) (p=0.17) in Group A and Group B respectively. The two schedules were comparable on using the Kruskal-Wallis H method for analysis. CONCLUSION: The schedule of hepatitis B vaccination at 0, 6 weeks and 9 months has the same seroefficacy as the currently recommended schedule of 0, 1 and 6 months.

Tubercular Osteomyelitis of Zygomatic bone.

A case of tubercular osteomyelitis of the zygomatic bone in a child is presented here because of its rarity and clinical interest. A high index of suspicion should be kept in mind to reach the diagnosis.

A study on the perinatal transmission of the hepatitis B virus.

The purpose of this study was to determine the age wise prevalence of Hepatitis B virus (HBV) in children under five years and to analyze the relative importance of horizontal or vertical transmission. This study included 400 children in the age group of less than five years attending the outpatient department of pediatrics with minor complaints. History of HBV immunization was taken as the exclusion criteria. All the samples were tested for Hepatitis B surface antigen (HBsAg) and anti HBs using commercial ELISA kits. Liver function tests were performed on all the HBsAg positive patients. Hepatits B nucleocapsid antigen (HBeAg) was detected in few HBsAg positive mothers. Overall HBsAg positivity in children below five years was 2.25%. There was no statistically significant difference in HBsAg positivity in the different age groups by chi square test. HBsAg positivity in mothers was 4.25%. However only in three cases the pair of mother and child were both positive for HBsAg. The mean anti HBs positivity in children was 23.75%. There was no statistically significant difference in the anti HBs positivity in different age groups of children. The observation that there is no statistically significant difference in the prevalence of HBV infection (HBsAg and HBs) amongst different age groups of children below five years signifies that a large proportion of HBV infection in children of this age is acquired via vertical transmission. It is also indicated that this mode of disease transmission is responsible for the majority of chronic carriers. Universal immunization of all infants is desirable to decrease the carrier pool and it is inferred from the present study that Hepatitis B immunization should begin at birth to have greater impact.

Anti-HCV seropositivity among multiple transfused patients with beta thalassaemia.

Hepatitis C virus is considered to be the main aetiological agent responsible for the occurrence of post-transfusion hepatitis. Patients with thalassaemia acquire hepatitis most often from viruses contracted through blood transfusions. The present study was undertaken to evaluate the prevalence of hepatitis C virus (HCV) in thalassaemic patients with multiple blood transfusions. The association of HCV seropositivity with number of blood transfusions and liver enzyme profile was also analysed. The study group consisted of fifty patients (40 males and 10 females) attending the thalassaemic unit of Lok Nayak Hospital, a tertiary care hospital at Delhi, within the age group of 1-25 years. Thirty patients (60%) were found to be seropositive for HCV antibodies while one patient (2%) was co-infected with HCV antibodies and hepatitis B surface antigen. Study of liver enzyme profile showed aspartate aminotransferase levels to be significantly higher, although the level of serum alanine aminotransferase, alkaline phosphatase, total protein, bilirubin and albumin were not significantly altered in these patients. It is inferred from this study that 60% of the thalassaemics were infected with HCV and this was directly related to the number of blood transfusions received by them. The regularised national blood policy followed by blood banks for providing safe blood along with better screening method of donated blood in blood banks would bring down the incidence of hepatitis C in such high risk group.

Comparison of two blood collection techniques for measles antibody detection in children.

BACKGROUND & OBJECTIVES: Though several seroprevalence and seroconversion studies have been conducted for measles using the filter paper haemagglutination inhibition (FPHAI), very few studies have compared the conventional serum HAI with the FPHAI. The present study was aimed at the evaluation of whole blood samples on filter paper as an alternative to serum specimens for detection of antibodies to measles virus. METHODS: Serum and whole blood samples soaked on filter paper were collected from 165 randomly selected healthy children in the age groups of 1-2 yr. HAI test was performed on both sets of samples and the results compared. RESULTS: Ninety samples that gave a titre of 8 or more by serum HAI also had titres of > or = 8 by FPHAI showing 100 per cent agreement between the two assays. Seventeen samples that had a serum HAI titre of 2 and 4 were missed by FPHAI due to the starting dilution of 8 of the latter. However, FPHAI gave no false positive results compared with serum HAI. INTERPRETATION & CONCLUSION: Collection of the FP samples by finger prick is more acceptable and requires less expertise than venepuncture. Thus, in spite of the small percentage of missed cases by the filter paper method, this technique of sampling was found to be a convenient and reliable alternative to venepuncture, for detection of measles virus antibody especially in large scale seroepidemiological studies.