Typing of rabies virus isolates by DNA enzyme immunoassay

Sabouraud, A.; Smith, Jean S.; Orciari, Lillian A.; Mattos, Carlos de Meira; Mattos, Cecilia De; Rohde, Rodney E.

doi:10.1016/s1386-6532(98)00006-7

Cited by 10 publications

(3 citation statements)

References 12 publications

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“…Newer detection assays have been developed that allow more rapid evaluation of PCR results (3,4,13,25,26,33,43). We developed a DEIA to be used in conjunction with the NLV and HAV RT-PCRs.…”

Section: Discussionmentioning

confidence: 99%

Development of a Reverse Transcription-PCR–DNA Enzyme Immunoassay for Detection of “Norwalk-Like” Viruses and Hepatitis A Virus in Stool and Shellfish

Schwab¹,

Neill²,

Guyader

et al. 2001

Appl Environ Microbiol

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Outbreaks of food-and waterborne gastroenteritis are being increasingly reported throughout the world. The analysis of environmental samples by newer diagnostic techniques such as reverse transcription-PCR (RT-PCR) amplification of nucleic acid has begun to identify human enteric viruses (predominantly "Norwalklike" viruses [NLVs]) as the cause of many of these outbreaks. To streamline NLV detection from environmental samples such as shellfish, we have developed an RT-PCR-oligoprobe amplification and detection method using several new procedures that enable confirmed RT-PCR amplification and product detection in 1 day. The new steps include replacing reverse transcriptase and Taq polymerase with rTth polymerase, a heat-stable enzyme that functions as both a reverse transcriptase and DNA polymerase, in a single-tube, single-buffer, elevated temperature reaction. An internal standard Norwalk virus (NV) RNA control is added to each RT-PCR to identify sample inhibition, and thermolabile uracil N-glycosylase is incorporated into the reaction to prevent PCR product carryover contamination. Finally, RT-PCR-generated amplicons are detected in microtiter wells using virus-specific biotinylated oligoprobes in an enzyme-linked immunosorbent assaybased format. The DNA enzyme immunoassay is based on the capture of PCR product by biotinylated probes fixed onto individual streptavidin-coated wells. Using this method, low levels of NV were detected in stool and both NLV and hepatitis A virus were detected in bivalve mollusks following bioaccumulation. The method also successfully detected NLV in oysters implicated in an outbreak of NLV gastroenteritis. This method dramatically decreases the time needed for analysis and is amenable to automation.

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Section: Discussionmentioning

confidence: 99%

Development of a Reverse Transcription-PCR–DNA Enzyme Immunoassay for Detection of “Norwalk-Like” Viruses and Hepatitis A Virus in Stool and Shellfish

Schwab¹,

Neill²,

Guyader

et al. 2001

Appl Environ Microbiol

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show abstract

“…• Is there a way to vaccinate wildlife using a recombinant vaccine dropped from aircraft? [3][4][5][6][7][8][9][10][11][12][13][14] • Can scientists actually determine the origin of a rabies virus variant by way of genetic testing and methodologies? [3][4][5][6][7][8][9][10][11][12][13][14] • Are there special government laboratories that conduct specialized rabies testing?…”

Section: Introductionmentioning

confidence: 99%

“…[3][4][5][6][7][8][9][10][11][12][13][14] • Can scientists actually determine the origin of a rabies virus variant by way of genetic testing and methodologies? [3][4][5][6][7][8][9][10][11][12][13][14] • Are there special government laboratories that conduct specialized rabies testing? [3][4][5][6][7][8][9][10][11][12][13][14] • Can international public health teams actually halt and remove a moving viral epizootic outbreak in a geographical area?…”

Section: Introductionmentioning

confidence: 99%