Whole genome analysis of Human Mastadenovirus D causing Keratoconjunctivitis in India - A multicentre study.

INTRODUCTION: Human mastadenovirus (HAdV) types 8, 37, 64 have been considered the major contributors in Epidemic keratoconjunctivitis (EKC) epidemics, but recent surveillance data have shown the involvement of emerging recombinants, including HAdV-53, HAdV-54, and HAdV-56. In our initial work, positive samples for adenovirus revealed that our strains were closer to HAdV-54 than HAdV-8. Hence, the current study aimed to use whole genome technology to identify the HAdV strain correctly. METHODOLOGY: Oxford Nanopore technique was used, wherein a Targeted sequencing approach using long-range PCR amplification was performed. Primers were designed using HAdV-54 (AB448770.2) and HAdV-8 (AB897885.1) as reference sequences. Amplicons were sequenced on the GridION sequencer. Sequences were annotated using Gatu software, and similarities with standard reference sequence was calculated using Bioedit software. The phylogenetic tree was built after alignment in MEGA v7.0 using Neighbour joining method for each of the genes: Penton, Hexon, and Fiber. The effect of novel amino acid changes was evaluated using the PROVEAN tool. The Recombination Detection Program (RDP) package Beta 4.1 was used to identify recombinant sequences. RESULTS: Of the five samples sequenced, OL450401, OL540403, and OL540406 showed nucleotide similarity to HAdV-54 in the penton region. Additionally, OL450401 showed a statistically significant recombination event with HAdV-54 as minor and HAdV-8 as major parents. This was further supported by phylogenetic analysis as well. CONCLUSIONS: In the present study, we have found evidence of a shift from HAdV-8 towards HAdV-54, thus stressing the need for surveillance of HAdVs and to stay updated on the rise of new recombinants.

Epidemiology of Keratoconjunctivitis Across India from 2017 to 2019: A Multicentric Hospital-Based Study.

PURPOSE: Conjunctivitis is one of the most common ocular conditions in clinical practice. Human adenoviruses have been the common causative agents known to cause epidemic kerato-conjunctivitis (EKC) in India from 1996 to 2019 with a positivity range of 13.8%-65.2%. The current study was initiated to throw light on the distribution of keratoconjunctivitis causing agents across India covering a span of 3 years. METHODS: A total of 709 swabs were collected from patients in viral transport medium (VTM), and real-time PCR was done to identify agents including Adenovirus (HAdV), Enterovirus, HSV, and Chlamydia. RESULTS: 47.8% of the samples were positive for HAdV followed by HSV (3.4%), Enterovirus (2.7%), and Chlamydia (0.6%). Overall, 386 people (54.4%) tested positive for one of these infections, with Chandigarh (88.4%) and Port Blair (71.7%) showing higher positivity rate. Pre-auricular lymphadenopathy and follicles were significantly associated with increased risk of conjunctivitis. CONCLUSION: Epidemiology of keratoconjunctivitis in the current study revealed HAdV to be predominant causative agent. Knowledge gained in such epidemiological studies guide us in outbreak expectations, limit antibiotic over-prescription, and enhance disease prevention.

A study on viral haemorrhagic fever due to dengue, chikungunya and Crimean Congo haemorrhagic fever virus among patients attending tertiary care hospital in North East India.

PURPOSE: The present study was undertaken with the objective to study the common etiology of Viral Haemorrhagic Fever (VHF) among patients attending tertiary health care centre in NE India and also to study the clinico-demographic profile of such patients. The agents of VHF included in the study were dengue, chikungunya and Crimean Congo haemorrhagic fever (CCHF) virus. The inclusion of CCHF was based on evidence of seroprevalence in livestock (bovine, sheep and goat) in various North Eastern states. MATERIALS AND METHODS: Serum samples were collected from 51 suspected VHF patients. MAC-ELISA was done to detect dengue and chikungunya specific IgM antibody. The samples were also tested by real-time RT-PCR for detection of dengue, chikungunya and CCHF specific nucleic acid. The laboratory and clinico-demographic profile of these patients were noted in detail. RESULTS: Serum samples of 16 of 51 suspected cases were confirmed to be suffering from VHF. Among these confirmed cases, 12 were diagnosed with dengue haemorrhagic fever, one was diagnosed with chikungunya and three were diagnosed with dengue-chikungunya co-infection. Based on severity, DHF was further classified into- DHF I- (4,26.6%), DHF II (6,40%), DHF III (3,20%) and DHF IV (2,13.3%). There was no CCHFV infection detected in our study. Retro-orbital pain (P â€‹= â€‹0.02) and haematocrit level (P â€‹= â€‹0.03) were found to be statistically significant. CONCLUSIONS: This study reiterates the fact that CCHF virus infection is still probably absent in human population of NE India and haemorrhagic symptoms, though rare maybe one of the atypical manifestations of chikungunya infection.

Ecology of Bloodstream Infections and Temporal Trends of Their Antibiograms with Respect to Source and Duration of Incubation: A 5-Year Retrospective Observational Analysis.

Purpose Blood is one of the most important connective tissues of human body. Bloodstream infection can range from inapparent bacteremia till fulminant septic shock with high mortality. Presence of microbes in blood whether continuously, intermittently, or transiently is a grave risk to every organ of body. Culture of blood is a vital tool to diagnose such infections. Drug susceptibility patterns help in rationalizing therapy. Objective The aim of the study is to perform bacteriological analysis and assess drug sensitivity patterns of blood culture isolates and compare in light of other associated variables. Design Retrospective observational study was conducted from January 2009 to December 2013 at a tertiary care hospital at Shillong, India. Blood samples were collected with aseptic guidelines and cultured for 7 days. Growths were identified by standard biochemical tests and subjected to sensitivity testing according to Modified Kirby Bauer disk diffusion method. Data for source of blood collection and duration of incubation were noted and compared. Results A total of 658 (11.2%) pathogens were isolated from 5,867 bacteremia-suspected patient blood specimens. Contamination was observed at the rate of 1.21%. Gram-negative organisms were the predominant pathogens recovered, Klebsiella pneumoniae being the most common. No significant difference was observed between the number of organisms isolated within or beyond 48 hours. Acinetobacter baumannii and K. pneumoniae have significantly higher chances ( p < 0.05) of isolation from central line catheters compared with peripheral venipuncture. Conclusion Successful treatment of sepsis depends on early diagnosis and proper antimicrobial therapy. Local knowledge of bacteriological profile and antimicrobial sensitivity patterns helps rationalize empiric treatment strategies.

Rotavirus associated acute diarrhoea in hospitalized children in Dibrugarh, north-east India.

Faecal specimens from 202 children below 5 years with acute diarrhoea hospitalized in Assam Medical College from April, 1999 to March, 2000 were examined in Regional Medical Research Centre (ICMR), Dibrugarh to know the prevalence of rotavirus diarrhoea and molecular pattern of viral strains from different localities of Dibrugarh using double antibody sandwich ELISA and SDS-PAGE analysis. Human group A rotaviruses were detected in 47 (23.27%) specimens and 33 of 41 (80.49%) positive specimens were electropherotyped where 16 were "long" (48.48%) and 17 "short" (51.52%) types. Rotavirus diarrhoea was significantly high (p<0.01) in children between 11 to 20 months (37.75%). Children from families of upper middle socioeconomic status (61.59%) suffer most (p<0.001). Peak incidence of rotavirus diarrhoea was in winter (38.37%) and showed inverse relation with temperature, humidity and rainfall. Besides diarrhoea, vomiting was a significant clinical manifestation. "Short" electropherotype were common during winter months and in tea garden localities.