Molecular characterization of influenza A(H1N1)pdm09 viruses circulating at various geographical locations in India, 2017

Background & objectives: Influenza virological surveillance is an essential tool for the early detection of novel genetic variants of epidemiologic and clinical significance. This study was aimed to genetically characterize A(H1N1)pdm09 virus circulating in 2017 and to compare it with the global data. Methods: The regional/State Viral Research and Diagnostic Laboratories (VRDLs) provided influenza diagnosis for referred clinical samples and shared influenza A(H1N1)pdm09 positives with the Indian Council of Medical Research-National Institute of Virology (ICMR-NIV), Pune, India, for hemagglutinin (HA) gene phylogenetic analysis. Sites at Manipal, Jaipur and Dibrugarh performed the sequencing and shared the sequence data for analysis. The antiviral susceptibility of influenza viruses was assessed for known molecular marker H275Y at the ICMR-NIV, Pune. Results: All the eight VRDLs had well-established influenza diagnostic facilities and showed increased activity of influenza A(H1N1)pdm09 during 2017. Phylogenetic analysis showed that the viruses from the different regions of the country were similar to A/Michigan/45/2015 strain which was the 2017-2018 recommended vaccine strain and were clustered with the globally circulating clade 6B.1 with signature mutations S84N, S162N and I216T. The clade 6B.1 showed further subgrouping with additional mutations S74R, S164T and I295V; however, there was no significant association between the presence of these mutations and severity of disease due to influenza. All the study viruses were sensitive to oseltamivir. Interpretation & conclusions: During the study period, all the study sites reported globally circulating A/Michigan/45/2015 vaccine strain of influenza A(H1N1)pdm09 viruses and remained sensitive to oseltamivir. Further genetic and antigenic characterization of influenza viruses is recommended to address public health concerns.

Geographical distribution of primary & secondary dengue cases in India – 2017: A cross-sectional multicentric study

Background & objectives: Dengue virus infection is endemic in India with all the four serotypes of dengue virus in circulation. This study was aimed to determine the geographic distribution of the primary and secondary dengue cases in India. Methods: A multicentre cross-sectional study was conducted at Department of Health Research / Indian Council of Medical Research (DHR)/(ICMR) viral research and diagnostic laboratories (VRDLs) and selected ICMR institutes located in India. Only laboratory-confirmed dengue cases with date of onset of illness less than or equal to seven days were included between September and October 2017. Dengue NS1 antigen ELISA and anti-dengue IgM capture ELISA were used to diagnose dengue cases while anti-dengue IgG capture ELISA was used for identifying the secondary dengue cases. Results: Of the 1372 dengue cases, 897 (65%) were classified as primary dengue and 475 (35%) as secondary dengue cases. However, the proportion varied widely geographically, with Theni, Tamil Nadu; Tirupati, Andhra Pradesh and Udupi-Manipal, Karnataka reporting more than 65 per cent secondary dengue cases while Srinagar, Jammu and Kashmir reporting as low as 10 per cent of the same. The median age of primary dengue cases was 25 yr [interquartile range (IQR 17-35] while that of secondary dengue cases was 23 yr (IQR 13.5-34). Secondary dengue was around 50 per cent among the children belonging to the age group 6-10 yr while it ranged between 20-43 per cent among other age groups. Interpretation & conclusions: Our findings showed a wide geographical variation in the distribution of primary and secondary dengue cases in India. It would prove beneficial to include primary and secondary dengue differentiation protocol in the national dengue surveillance programme.

Profile of respiratory pathogens causing acute respiratory infections in hospitalised children at Rajasthan a 4 year's study

Introduction: Various pathogens cause respiratory tract infections in children of <5 years of age causing severe morbidity and mortality. The profile of causative agents varies from place to place. Aims: The objectives of our study were to detect the profile and trends of respiratory pathogens causing acute respiratory tract infection in children using a custom multiplex real-time polymerase chain reaction (RT-PCR) and to develop a diagnostic algorithm. Materials and Methods: A total of 997 children with clinical manifestations of respiratory infections were included in the study. Their nasopharyngeal aspirate and throat swab samples were subjected to nucleic acid extraction followed by multiplex RT-PCR for eighteen viruses and six bacteria. Statistical Analysis Used: Chi-square test was employed to study the P value of different viruses and bacteria. Results: A total of 765 (76.73%) samples were found to be positive for one of the respiratory pathogens. Viruses were detected in 598 (59.98%) and bacteria in 167 (41.85%) samples, respectively. The prevalence of single and co-infections among viruses and bacteria were 77.76% and 22.24%, 81.44% and 18.56% each, respectively. Respiratory syncytial virus (RSV) A/B and Streptococcus pneumoniae were the most predominant pathogens detected in the study and were associated with lower respiratory tract infections. Conclusion: RSV and S. pneumoniae were the most common pathogens detected, higher prevalence was observed in children <1 year of age. Viruses were predominant during winter months. The study helped to prepare diagnostic algorithm which will help in reducing diagnostic costs. However, further studies are required to assess whether viruses are bystander or real pathogens and include larger panel of bacteria and viruses for diagnosis.

Molecular characterization of hepatitis A virus from children hospitalized at a tertiary care centre in northwest India

Background & objectives: Hepatitis A virus (HAV) infection is a major cause of childhood hepatitis, prevalent worldwide. HAV is classified into seven genotypes I-VII; genotypes III and I are the most common among humans. The present work was carried out to identify the genotypes prevalent in children suspected to have acute viral hepatitis (AVH), hospitalized at a tertiary care centre in northwest India. Methods: A total of 1269 blood samples from children (0-15 yr of age) clinically suspected of viral hepatitis were screened for anti-HAV IgM. Acute phase serum was processed for RNA extraction and amplified by nested polymerase chain reaction (PCR) followed by sequencing of representative samples. Results: Among the 1269 samples tested, 642 (50.59%) were positive for anti-HAV IgM; among the positive samples, 171 patients having a history of less than seven days were tested by PCR, of whom 141 (82.45%) were found to be PCR positive. Nucleotide sequencing of a representative 44 samples showed high homology; all the samples were found to be of genotype IIIA. Interpretation & conclusions: Hepatitis A was prevalent during July to September and in predominantly children less than five years age. Only genotype IIIA was detected in all the samples.

A comparative study for the detection of Mycobacteria by BACTEC MGIT 960, Lowenstein Jensen media and direct AFB smear examination.

PURPOSE: To compare BACTEC MGIT 960 (M960) with conventional culture on Lowenstein Jensen (LJ) media and direct acid fast bacilli (AFB) smear examination for the detection of Mycobacteria in clinical samples obtained from suspected cases of pulmonary and extra pulmonary tuberculosis (TB). METHODS: A total of 500 samples were processed for direct AFB smear examination, and culture on M960 and LJ media. RESULTS: Two hundred fifty-eight out of 500 (51.6%) isolates of Mycobacteria were obtained by combined use of the two culture methods. Two hundred and fifty-three (50.6%) were positive in culture by M960 and LJ media and 28% (140/500) by direct AFB smear examination. The positivity rate of M960 system alone was 34.10% (88/258) and of LJ alone was 1.93% (5/258). Average time to detect growth (TTD) was 9.66 days by M960 and 28.81 days by LJ. CONCLUSIONS: M960 system is a rapid and sensitive method for early diagnosis of pulmonary and extrapulmonary TB. But for maximum recovery of Mycobacteria , a combination of both M960 and LJ media should be used.

Molecular typing of Mycobacterium tuberculosis isolates from different parts of India based on IS6110 element polymorphism using RFLP analysis.

BACKGROUND & OBJECTIVE: IS 6110 based typing remains the internationally accepted standard and continues to provide new insights into the epidemiology of Mycobacterium tuberculosis. The aim of the study was to characterize M. tuberculosis isolates obtained from different parts of India based on IS6110 element polymorphism using restriction fragment length polymorphism (RFLP) analysis. METHODS: RFLP was analyzed among 308 isolates of M. tuberculosis deposited in the Mycobacterial Repository Centre, Agra, from different parts of India. DNAs isolated from these strains were restricted with Pvu II, transferred on to nylon membrane and hybridized with a PCR amplified DIG-labeled 245 bp IS6110 probe. RESULTS: Based on the copy number, M. tuberculosis isolates were classified into four groups, (i) lacking IS6110 element; (ii) low copy number (1-2); (iii) intermediate copy number (3-5); and (iv) high copy number (6-19). Copy number higher than 19 however was not observed in any of the isolates studied. At the national level, 56 per cent of the isolates showed high copy number of IS6110, 13 per cent showed intermediate copy number, 20 per cent showed low copy number, whereas 11 per cent isolates lacked IS6110 element. At the regional level, there was not much difference in the RFLP profiles of isolates (IS6110 copy numbers/patterns) from different parts of the country. INTERPRETATION & CONCLUSION: IS6110 DNA based fingerprinting could be a potentially useful tool for investigating the epidemiology of tuberculosis in India.

Drug susceptibility profiles of mycobacterium tuberculosis isolates at Jaipur.

PURPOSE: To find prevalence of drug resistance in Mycobacterium tuberculosis isolated from patients attending SMS Medical College, Jaipur during 1997-99. METHODS: Sputum samples from 164 patients with pulmonary tuberculosis were processed and cultured on Lowenstein Jensen medium and M.tuberculosis isolates were tested for drug sensitivity. RESULTS: M. tuberculosis was isolated in 122/164 (74.3%) samples and comprised 97.6% (122/125) of mycobacterial isolates. There were only three isolates of nontuberculous mycobacteria -one each of M.kansasii, M.avium and M.fortutium. Primary drug resistance in M.tuberculosis was estimated to be 3/44 (6.8%) to rifampicin, 6/44 (13.6%) to isoniazid and 2 strains (4.5%) were multi drug resistant i.e. resistant to both rifampicin and isoniazid. Among the isolates from cases with previous history of treatment of varying duration (acquired drug resistance) resistance to rifampicin was estimated to be 28.2% and for isoniazid to be 39.7%. 24.3% strains of these drug resistant isolates were multi drug resistant. CONCLUSIONS: While this information may not reflect true prevalence of drug resistance in the region this may help in further planning long term surveillance studies to know the trend of drug resistance in this area.

Port site metastasis subsequent to laparoscopy for chronic pelvic pain.

We present an interesting case of port site metastasis is a menopausal women subsequent to diagnostic laparoscopy undertaken for chronic pelvic pain, which later proved early ovarian malignancy as the source of primary. While cases of port site metastasis have mostly occurred after extensive disease the possbility of such complication should be in mind at laparoscopy of early cancer.

Polymyositis--an unusual presentation of ovarian malignancy: a case report.

A unique case of carcinoma of ovary with an unusual presentation is reported here. Her presenting complaint was in no way related to ovarian malignancy. She presented to the neurologist with the features suggestive of polymyositis. She did not respond satisfactorily to the conventional treatment by steroids and on investigation, was found to have an underlying advanced ovarian malignancy.

An unusual foreign body in tracheobronchial tree: removed by fiberoptic bronchoscope.

Tracheostomy is a life saving procedure and many a times patients are discharged with permanent tracheostomy tubes. Rarely, a tracheostomy tube may break and enter the tracheobronchial tree. We present a case of an adult female in whom the broken tracheostomy tube entered the right main bronchus and was removed by fiberoptic bronchoscopy under topical anaesthesia.

Liquid membrane phenomenon in the actions of digitalis.

The liquid membrane phenomenon in the actions of digitalis glycosides (digitoxin, digoxin and ouabain) has been studied. Formation of liquid membranes, in series with a supporting membrane, by digitalis alone and by digitalis in association with lecithin and cholesterol has been demonstrated. The results obtained on the transport of relevant permeants, viz. sodium, potassium and calcium ions and dopamine, adrenaline, noradrenaline and serotonin, in the presence of the liquid membrane generated by digitalis in association with lecithin and cholesterol indicate that the liquid membrane barrier to transport may have a relevance with the biological actions of digitalis.