First isolation and characterization of SARS-CoV-2 from COVID-19 patient of North East India.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) responsible for the current pandemic has resulted in over 5 million deaths globally. More than a year has passed, still SARS-CoV-2 panic the public life. Virus isolation is of paramount importance for development of vaccines, in-vitro screening of antiviral compounds, pathogenesis studies, etc., Many cell lines were studied for amplification and replication of SARS-CoV-2 and Vero cells were found to be ideal cell lines for isolation. In May 2020, ICMR-Regional Medical Research Centre, NE region, India, successfully established the SARS-CoV-2 culture system in Vero CCL-81 cell lines. Phylogenetic analyses of the whole genome sequences of the SARS-CoV-2 isolate (EPI_ISL_2501532 | 2020-05-19) showed monophyletic clade G and lineage B.1.1.

Prevalence of campylobacter enteritis in children under 5 years hospitalised for diarrhoea in two cities of Northeast India.

Background: Campylobacter enteritis is the major cause of bacterial gastroenteritis worldwide. In recent years, there has been a rise in global incidence of campylobacteriosis. There are no available data on prevalence of Campylobacter diarrhoea from Northeast India. Materials and Methods: The study investigated archival stool samples collected between 2014 and 2016 from two hospitals of Northeast India. A total of 407 archival stool samples from cases of diarrhoea under 5 years of age were screened for Campylobacter spp. using commercial probe-based real-time polymerase chain reaction assay. Results: Campylobacter spp. was detected in overall 10.1% (41/407; 95% confidence interval: 7.4%-13.3%) in children under 5 years hospitalised for diarrhoea. The prevalence was significantly higher from Dibrugarh, Assam, compared to Dimapur, i.e., 13.4% (27/201) versus 6.8% (14/206), respectively (P = 0.02). Campylobacter detection was highest in the month of June and July compared to December and January (20%-18.8% vs. 8.9%-6.2%, respectively). Further, Campylobacter infection was higher in the age group below 24 months (11.7%) compared to above 24 months (7.0%). Campylobacter jejuni was detected in 80.5% of the positive cases. Conclusion: The present study reveals that Campylobacter infection is endemic in the studied regions of Northeast India and microbiological laboratories of the region should actively pursue the isolation or detection of Campylobacter spp. in cases of diarrhoea in routine stool cultures.

Genetic variations of the Hemagglutinin gene of Pandemic Influenza A (H1N1) viruses in Assam, India during 2016.

Since its emergence in 2009, Influenza A/H1N1pdm09 virus has evolved continuously. Marked genetic variations have occurred in the HA1 domain of the hemagglutinin gene causing the emergence of new variants. The present study genetically characterized the hemagglutinin (HA) gene of Influenza A/H1N1pdm09 strains from Assam circulating in 2016 that caused a mild outbreak without any reported mortality. Sequence analysis of the HA gene of 20 positive Assam/H1N1pdm09 strains revealed 3 mutations (K180Q, S202T, S220T) at the antigenic sites along with several other reported mutations which are in close proximity to the antigenic sites and therefore might affect the viral antigenicity. Phylogenetically, the Assam/H1N1pdm09 strains clustered into genogroup 6B. These genetic variations highlight the importance of continuous surveillance and characterization of Influenza A/H1N1pdm09 virus activity to track the genetic makeup and diversification that may affect the behavior of the virus.

Molecular characterisation and phylogenetic analysis of dengue outbreak in Pasighat, Arunachal Pradesh, Northeast India.

BACKGROUND AND OBJECTIVES: Dengue is one of the most prevalent arboviral diseases in the world with 390 million dengue infections per year. In this study, we report the molecular characterisation of dengue outbreak in Pasighat, Arunachal Pradesh, Northeast India during 2015. SUBJECTS AND METHODS: : A total of 613 dengue-suspected cases were screened for dengue virus by dengue NS1 Ag and anti-dengue IgM antibody depending on the duration of sample collection and onset of symptom. Further, molecular characterisation was done by amplifying the C-PrM region by real-time polymerase chain reaction followed by phylogenetic analysis. RESULTS: Molecular characterisation revealed that the dengue outbreak was predominantly due to dengue virus serotype-1 (DENV-1) (90.9%) while DENV-2 was detected in 7.5% of samples. Co-infection of DENV-1 and DENV-2 was detected in one case. Phylogenetic analysis of the DENV-1 strains with the prototype revealed that the DENV-1 strains were grouped within genotype III. Similarly, DENV-2 strains were clustered within genotype IV. The study revealed a change in the predominant serotype in recent years with DENV-3 in 2012 to DENV-1, 2, 3 and 4 in 2014 to DENV-1 in 2015 in the study region. A unique L24M mutation was observed in the DENV-1 strains of Arunachal Pradesh which was absent in all the circulating strains in India except one strain from the state of Kerala in South India. Marked variation within the DENV-2 strains was observed at A102V and I163V in one strain similar to earlier circulating isolates in India. CONCLUSIONS: The present study reveals a shift in the serotype dominance in the study region. As serotype shifts and secondary infection with a heterologous DENV serotype are frequently associated with disease severity, there is an urgent need for sustained monitoring of the circulating serotypes and enhanced surveillance operations, especially in the monsoon and post-monsoon periods to prevent large-scale, severe dengue outbreaks in this region.

Seroprevalence of Toxoplasma gondii Among Pregnant Women in Northeast India.

OBJECTIVE: Toxoplasma gondii infection is primarily asymptomatic and one third of world's population is estimated to be infected by this protozoan parasite. This study was carried out to determine the seroprevalence of T. gondii infection among pregnant women from north east India, where data on this important parasitic infection is scanty. METHODS: A total of 1141 serum archival samples collected from antenatal clinic attendees in 2007-09, were screened for T. gondii IgG by ELISA and analyzed with their socio demographic information. RESULTS: The median age of the subjects were 25 years with an overall IgG seroprevalence of 48% (95% CI=45% to 51%). Seroprevalence was significantly associated with geographical location (p=0.000), among Mongoloids compared to Caucasoid (p=0.005), regular employees (p=0.003) or unskilled labors (p=0.04) compared to housewives, high or middle income group (p=0.003) compared to low income group and with increasing gravida (p=0.04). However on logistic regression analysis, only significant association was with geographical location (p=0.000) and occupation (unskilled labor) (p=0.04). Highest prevalence of 66.7% was noted among subjects with history of blood transfusion and lowest among Rh negative blood group (14.3%). CONCLUSIONS: T. gondii infection prevalence is high among pregnant women living in hilly states of northeast India, which warrants a detail investigation on congenital toxoplasmosis as well as its economic impact.

Seroprevalence of Scrub Typhus Infection in Arunachal Pradesh, India.

INTRODUCTION: Scrub typhus is a major reason for febrile illness, caused by a bacterium Orientia tsutsugamushi, a rickettsial pathogen. Few outbreaks of scrub typhus have been reported from Arunachal Pradesh in recent past. However, there is lack of seroprevalence data from the region. In this regard, this study was undertaken using archival serum sample available from seven districts of Arunachal Pradesh. METHODOLOGY: This serological study was conducted in Regional Medical Research Center for NE Region, Dibrugarh. Reactivity to IgG class of antibodies against scrub typhus was done using Scrub typhus detect IgG ELISA kit as per manufacturer's protocol. RESULT: Seroprevalence of scrub typhus in seven districts of Arunachal Pradesh was found to be 40% (120/300). The age-specific scrub typhus seroprevalence rose steadily from 5.6% in children <10 years of age to 61.8% in persons aged ≥40 years (p = 0.0001). Prevalence is lowest in Papumpare (25.9%) and highest in East Siang (72.5%) (p = 0.0001). The seroprevalence in males and females was very similar, however, the female prevalence increases from age group ≥30 years (p = 0.053). Moreover, among the farmers, the seroprevalence is higher (58.3%) (p = 0.0001). CONCLUSIONS: As clinical symptoms overlap with other viral/bacterial infections, scrub typhus infection should be considered in differential diagnosis of any acute febrile illness in this part of the country. In view of the high prevalence, empirical therapy of doxycycline/azithromycin may be done in cases of undiagnosed fever. Active surveillance has to be done to understand exact magnitude, epidemiological aspects, and distribution of vector and disease of this reemerging neglected tropical disease.

Expression of SPARC like protein 1 (SPARCL1), extracellular matrix-associated protein is down regulated in gastric adenocarcinoma.

BACKGROUND: SPARC-like protein 1 (SPARCL1/Hevin), a member of the SPARC family is defined by the presence of a highly acidic domain-I, a follistatin-like domain, and an extracellular calcium (EC) binding domain. SPARCL1 has been shown to be down-regulated in many types of cancer and may serve as a negative regulator of cell growth and proliferation. METHODS: Both tumor and adjacent normal tissue were collected from patients with gastric adenocarcinoma. Monoclonal antibody developed against recombinant SPARCL1 was used to analyze the expression of SPARCL1 by immunohisto chemical and western blotting (WB) analysis. RESULTS: The expression of SPARCL1 was found to be significantly lower or negligible in gastric adenocarcinoma tissues in nearly all of the cases in comparison with adjacent normal tissue. This comparison was found to be independent of the patient's age, sex, and stage of cancer. CONCLUSIONS: We postulate that down regulation of SPARCL1 may be related to inactivation of its tumor suppressor functions and might play an important role in the development of gastric adenocarcinoma.

Co-infection of scrub typhus and leptospirosis in patients with pyrexia of unknown origin in Longding district of Arunachal Pradesh in 2013.

BACKGROUND: Scrub typhus and leptospirosis are bacterial zoonotic disease causing high morbidity and mortality. The seasonal outbreak of pyrexia is common in Arunachal Pradesh (AP); many times the disease remains undiagnosed. OBJECTIVE: An outbreak of pyrexia of unknown origin (PUO) occurred in Longding district of Arunachal Pradesh in 2013, with 108 deaths, which was investigated to elucidate the cause of illness. METHODOLOGY: Blood samples from the affected region with acute pyrexia were collected, and screened for the malaria parasite, scrub typhus IgM and leptospira IgM. RESULTS: Scrub typhus IgM was reactive in 97% (30/31), and 25% (8/31) cases were co-infected with leptospira. Incidentally, scrub typhus reactive (67%) and leptospira co-infection (62.7%) were higher in females. Record of previous 3 years (2011-2013) from Longding, Community Health Centre showed an increase in indoor pyrexia cases by 2-fold or more during October and November. CONCLUSION: The present study is the first report of co-infection of scrub typhus with leptospirosis from Northeast India. Medical officers in this region should take scrub typhus and leptospirosis in their differential diagnosis of patients with PUO for early diagnosis and effective treatment.

Gene Expression Profiling of Gastric Cancer.

Gastric cancer is the second leading cause of cancer death worldwide, both in men and women. A genomewide gene expression analysis was carried out to identify differentially expressed genes in gastric adenocarcinoma tissues as compared to adjacent normal tissues. We used Agilent's whole human genome oligonucleotide microarray platform representing ~41,000 genes to carry out gene expression analysis. Two-color microarray analysis was employed to directly compare the expression of genes between tumor and normal tissues. Through this approach, we identified several previously known candidate genes along with a number of novel candidate genes in gastric cancer. Testican-1 (SPOCK1) was one of the novel molecules that was 10-fold upregulated in tumors. Using tissue microarrays, we validated the expression of testican-1 by immunohistochemical staining. It was overexpressed in 56% (160/282) of the cases tested. Pathway analysis led to the identification of several networks in which SPOCK1 was among the topmost networks of interacting genes. By gene enrichment analysis, we identified several genes involved in cell adhesion and cell proliferation to be significantly upregulated while those corresponding to metabolic pathways were significantly downregulated. The differentially expressed genes identified in this study are candidate biomarkers for gastric adenoacarcinoma.