RT-PCR

Detection of Human Rotavirus by Reverse Transcription-Polymerase Chain Reaction (RT-PCR).

Abstract

Rotaviruses are routinely diagnosed by detection of rotavirus antigen in stools utilising an enzyme immunoassay (EIA). A perceptive procedure, like reverse transcription polymerase string of connections answer (RT-PCR), may disclose more rotaviruses, but the clinical significance of such outcome is not well established. To study if RT-PCR can notice more episodes of rotavirus-associated gas-troenteritis than EIA and to work out how rotavirus RT-PCR outcome might change efficacy investigation of a rotavirus vaccine test, in which the conclusion assess was rotavirus gas-troenteritis diagnosis with EIA.

RT-PCR directed for detection of rotaviruses in gas-troenteritis episodes came across in an efficacy test of rhesus-human reassortant rotavirus tetravalent (RRV-TV) vaccine, in a total of 2398 infants. During a follow-up, covering two rotavirus epidemic times of the year, 256 situations of rotavirus affiliated gastroenteritis were noticed by EIA; 226 were in the prime efficacy investigation time span that encompassed young children who had obtained three doses of vaccine or placebo.

With RT-PCR, 84 (33%) more situations of rotavirus gas-troenteritis were identified than with EIA, 65 of these were in the prime efficacy investigation period. Clinically, situations of rotavirus gas-troenteritis identified by RT-PCR were much milder (median severity tally 6 on a 20-point scale) than those identified by EIA (median tally 11), PB0.0001. RT-PCR disclosed proportionally more G2 and G4 rotaviruses than EIA. G1 rotaviruses noticed by RT-PCR were nearly identically split up between RRV-TV (25) vaccine and placebo (28) assemblies, while an clear-cut vaccine shielding effect was glimpsed in the circulation of G2 (one in the RRV-TV and eight in the placebo group) and G4 rotaviruses (six in the RRV-TV and 14 in the placebo group).

RT-PCR is a helpful device in the diagnosis of rotavirus gas-troenteritis, especially for situations affiliated with other than the epidemiologically superior G-type. Application of RT-PCR assists to the general appraisal of presentation of rotavirus vaccine.

Detection of Human Rotavirus by Reverse Transcription-Polymerase Chain Reaction (RT-PCR).

Introduction

Rotaviruses are the most important causative agents of severe acute gastroenteritis in young children world-wide, responsible for 600000800000 deaths annually (Ho et aI., 1988; Bern and Glass, 1994; Kapikian and Chanock, 1996). In developed countries, deaths are rare, but rotaviruses still cause a significant burden of disease, being associated with 30-60% of all hospitalizations due to acute gastroenteritis in children (Brandt et aI., 1983; Konno et aI., 1983; Glass et aI., 1996; Kapikian, 1996).

In our large efficacy trial of RR V-TV vaccine, the endpoint was rotavirus gastroenteritis diagnosed by EIA (Joensuu et aI., 1997). However, we retrospectively studied with R T - PCR, all stool specimens collected during the trial from episodes of acute gastroenteritis found rotavirus negative by EIA. The purposes of this study were: (1) to estimate how many cases of rotavirus gas-troenteritis might be missed using standard diagnostic methods; (2) to characterize virologically, by G-typing and clinically, cases of rotavirus gas-troenteritis detected only by R T - PCR in comparison with those detected by EIA; and (3) to describe how the results of RR V-TV vaccine trial ...