ABSTRACT
Background & objectives: Polio, measles, rubella, influenza and rotavirus surveillance programmes are of great public health importance globally. Virus isolation using cell culture is an integral part of such programmes. Possibility of unintended isolation of SARS-CoV-2 from clinical specimens processed in biosafety level-2 (BSL-2) laboratories during the above-mentioned surveillance programmes, cannot be ruled out. The present study was conducted to assess the susceptibility of different cell lines to SARS- CoV-2 used in these programmes. Methods: Replication of SARS-CoV-2 was studied in RD and L20B, Vero/hSLAM, MA-104 and Madin–Darby Canine Kidney (MDCK) cell lines, used for the isolation of polio, measles, rubella, rotavirus and influenza viruses, respectively. SARS-CoV-2 at 0.01 multiplicity of infection was inoculated and the viral growth was assessed by observation of cytopathic effects followed by real-time reverse transcription–polymerase chain reaction (qRT-PCR). Vero CCL-81 cell line was used as a positive control. Results: SARS-CoV-2 replicated in Vero/hSLAM, and MA-104 cells, whereas it did not replicate in L20B, RD and MDCK cells. Vero/hSLAM, and Vero CCL-81 showed rounding, degeneration and detachment of cells; MA-104 cells also showed syncytia formation. In qRT-PCR, Vero/hSLAM and MA-104 showed 106 and Vero CCL-81 showed 107 viral RNA copies per ?l. The 50 per cent tissue culture infectious dose titres of Vero/hSLAM, MA-104 and Vero CCL-81 were 105.54, 105.29 and 106.45/ml, respectively. Interpretation & conclusions: Replication of SARS-CoV-2 in Vero/hSLAM and MA-104 underscores the possibility of its unintended isolation during surveillance procedures aiming to isolate measles, rubella and rotavirus. This could result in accidental exposure to high titres of SARS-CoV-2, which can result in laboratory acquired infections and community risk, highlighting the need for revisiting biosafety measures in public health laboratories
ABSTRACT
Background: Down syndrome (DS) is a major cause of mental retardation of prenatal origin and has several associated co-morbidities involving cardiovascular system, respiratory, endocrine system, hematological, gastrointestinal, musculoskeletal, eye and ear defects, immunological changes and neurological system. This study was conducted to identify the common medical problems in children with Down syndrome and the morbidity associated with these conditions. The objective of the present study was to find out the occurrence of different medical problems in children with DS.Methods: 42 children with a phenotype of Down syndrome in the age group of 0-12 years attending the outpatient, inpatient and Down syndrome Clinic of the Institute of Child Health, Kottayam during the study period were included in the study by purposive sampling. Demographic details were entered, and Pediatric Clinical Examination was performed by the investigator himself to identify the medical problems. Old medical reports were reviewed, and data entered into a proforma and statistically analysed.Results: Out of the 42 children with DS, 22 were males. 15 (35.7%) were less than 1 year, 20 (48.3%) children 1-5 years and 7 (16.1%) children 5-12 years of age. Mean age of the study group was 1.78±0.51 years. Mean age of their mothers at the time of conception was 30.6±5.8 years. 26 (57%) children with Down syndrome had a medical problem during the neonatal period which required hospitalization. Almost all systems are affected and craniofacial features, developmental delay and hypotonia were universal. Various forms of congenital heart diseases were observed in 67% and hypothyroidism in 23.8%.Conclusions: Down syndrome is a common genetic disorder with multisystem involvement. Congenital heart diseases, hypothyroidism and recurrent respiratory infections were the common medical problems identified in this study.
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Background & objectives: The poliovirus serotype identification and intratypic differentiation by real-time reverse transcription-polymerase chain reaction (rRT-PCR) assay is suitable for serotype mixtures but not for intratypic mixtures of wild and vaccine poliovirus strains. This study was undertaken to develop wild poliovirus 1 and 3 (WPV1 and WPV3) specific rRT-PCR assays for use. Methods: Specific primers and probes for rRT-PCR were designed based on VP1 sequences of WPV1 and WPV3 isolated in India since 2000. The specificity of the rRT-PCR assays was evaluated using WPV1 and WPV3 of different genetic lineages, non-polio enteroviruses (NPEVs) and mixtures of wild/wild and wild/Sabin vaccine strains. The sensitivity of the assays was determined by testing serial 10-fold dilutions of wild poliovirus 1 and 3 stock suspensions of known titre. Results: No cross-reactivity with Sabin strains, intertypic wild poliovirus isolates or 27 types of NPEVs across all the four Enterovirus species was found for both the wild poliovirus 1 and 3 rRT-PCR assays. All WPV1 and WPV3 strains isolated since 2000 were successfully amplified. The rRT-PCR assays detected 104.40CCID50/ml of WPV1 and 104.00CCID50/ml of WPV3, respectively either as single isolate or mixture with Sabin vaccine strains or intertypic wild poliovirus. Interpretation & conclusions: rRT-PCR assays for WPV1 and WPV3 have been validated to detect all the genetic variations of the WPV1 and WPV3 isolated in India for the last decade. When used in combination with the current rRT-PCR assay testing was complete for confirmation of the presence of wild poliovirus in intratypic mixtures.
ABSTRACT
Introduction: Human obesity is strongly associated with cardiovascular disease. Cystatin C is a naturally occurring protease inhibitor and marker of cardiovascular disease. The atherogenic indices are used as an index for cardiac risk stratification. Objectives: To estimate the serum levels of Cystatin C in individuals with normal BMI, and obese, aged between 20-39 Yrs and to compare the levels of Cystatin C among these individuals and to correlate the levels of serum Cystatin C with atherogenic index of plasma and other indices. Methodology: The study population was taken from healthy volunteers of Mysore city, aged between 20-39 years of either sex. The study population was divided into 2 groups based on BMI. Each group contains sample size of 60. Fasting serum sample was analyzed for Total Cholesterol, TG, LDL-Cholesterol & HDL cholesterol by enzymatic method and serum Cystatin-C by immune-turbidimetric method using auto-analyser. Statistical Analysis: Analysis of Variance [ANOVA] was used to compare the serum levels of Cystatin C in the two groups. To correlate the serum Cystatin C with atherogenic indices for predicting the cardiovascular risk factors, Pearson’s correlation co-efficient was worked out. Results: The mean serum cystatin C levels in normal BMI group are 0.7±0.03 mg/L, and in Obese group 1.15±0.09 mg/(p value<0.001).In the study serum Cystatin C showed a positive correlation with serum triglycerides (r=0.7), Atherogenic index of plasma(AIP ) (r=0.80), TCHOL: HDL (Castelli’s Risk Index I) (r=0.71), HDL: LDL(Castelli’s Risk Index II) (r=0.70) respectively and Atherogenic coefficient (AC) {(NonHDLc)/HDLc}( r=0.60) and negative correlation with serum HDL(r=-0.52) Conclusion: Several indices had been derived from lipid profiles to establish an index for predicting the risk of having coronary event. The atherogenic index of plasma was strongly correlated with the Cystatin C, hence AIP can be used as better index for predicting the preclinical cardiovascular disease because of cost effectiveness in estimation of Cystatin C