Rotavirus Samples processing using Indigenously Developed Molecular Transport Medium (IMTM)

Abstract

Objective: To perform comparative diagnostic study with rotavirus infected stool samples for efficacy of
indigenously prepared molecular transport medium (IMTM).
Study Design: It was an experimental study.
Place and Duration of Study: The study was conducted at Department of Microbiology of University of Karachi,
Karachi Pakistan from June to July 2021.
Materials and Methods: The current study was conducted based on hospital based surveillance of rotavirus
gastroenteritis among children <5 years of age, stool samples were collected in commercially available viral
transport medium. Samples were known rotavirus positive /negative through PREMIER Rota clone Enzyme-
Linked Immunosorbent Assay (ELISA) kit diagnostic test. Total 64 stool samples were selected to evaluate the
efficacy of indigenous molecular transport medium (IMTM). Stool samples were spiked in IMTM and
commercial available viral transport medium (CVTM). RNA extraction and Real-time PCR were performed by
QIAmp and in house assay respectively as per manufacturer' instructions. Similar method of RNA extraction
and Real-Time PCR for both type of spiked samples was adopted.
Results: Comparative analysis of Cycle threshold (Ct) values of paired samples (IMTM vs CVTM) was performed.
Real-Time (RT-PCR) for rotavirus positive and negative results were consistent with each pair (p<0.0001).
Conclusion: The indigenously formulated IMTM found capable for stool sample collection, transportation and
processing for rotavirus detection through Real-time PCR.