Rapid screening test for the diagnosis of rotavirus infection

Abstract

The early diagnosis of human rotavirus infection is essential for effective patient management and infection control. We report here a rapid, easy-to-perform, and inexpensive test for rotavirus detection. The viral RNA is extracted directly from the stools and electrophoresed on 1% agarose gels. Currently available immunoassays for routine diagnostic purposes are directed at the common group A-specific antigen. As reports become available on human gastroenteritis caused by the atypical or novel rotaviruses, this technique presents an added advantage in that it can detect both group A and non-group A rotaviruses.

Comparison of agarose gel electrophoresis and RNA-PAGE for rapid detection of rotavirus from faecal samples

“…All the 11 bands of 11 segments of dsRNA were clearly visualised in all the concentrations of gel (Figure 1a-d). Similar results were also obtained by Chudzio et al (1989). In contrast, Pšikal et al (1991) obtained only eight and nine bands in 1.5% agarose gel.…”

“…Electrophoresis was performed as per the procedure described by Chudzio et al (1989) to determine the presence of rotavirus. Ready-to-use ethidium bromide (2-3 µl) for staining was added directly into the agarose (Sigma) during its preparation.…”

Detection of rotavirus and norovirus among children with acute gastroenteritis in Merida and Chihuahua cities, Mexico

“…18068). For RV detection, viral RNA was analyzed on a 7.5% PAGE and silver stained as described [13] and (Figure 1). HuNoV screening was performed by RT-PCR as previously described [14], using primers 289H and 289I [15] and 290YM [14] evolutionary history was inferred using the Neighbor-Joining method.…”

Ovine rotavirus strain LLR-85-based bovine rotavirus candidate vaccines: Construction, characterization and immunogenicity evaluation

“…RNA polyacrylamide gel electrophoresis (PAGE) was done as previously described (Chudzio et al, 1989). Briefly, the supernatant of the culture lysates were mixed with an equal volume of RNA extraction buffer (0.02 M Trishydrochloride [pH 7.4], 0.3 M NaCl, 0.01 M MgCl 2 , 0.1% sodium dodecyl sulfate, 0.005 M EDTA, 4% sucrose and 0.04% bromophenol blue), and each preparation was mixed with an one-tenth volume of phenol-chloroform (1:1) and vortexed for 30 s to yield a homogeneous suspension followed by spinning in a microcentrifuge at 12,000 Â g for 10 min.…”