Open AccessTranscriptional enhancement of RT‐PCR for rapid and sensitive detection of Noroviruses
Author(s) -
Jean Julie,
D'Souza Doris,
Jaykus LeeAnn
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00621-9
Subject(s) - nasba , amplicon , norovirus , microbiology and biotechnology , biology , polymerase chain reaction , reverse transcription polymerase chain reaction , reverse transcriptase , real time polymerase chain reaction , electrochemiluminescence , virology , detection limit , nucleic acid , virus , chemistry , rna , gene , messenger rna , chromatography , genetics
Previously reported nucleic acid sequence‐based amplification (NASBA) primers specific for the GII Noroviruses were adapted for reverse transcriptase‐polymerase chain reaction (RT‐PCR), and detection sensitivity was then enhanced by a subsequent in vitro transcription of the RT‐PCR amplicons. The NASBA‐derived primers performed comparably to other broadly reactive GII Norovirus primers with respect to detection limits (i.e. 1 RT‐PCR amplifiable unit (RT‐PCRU) per reaction). Detection limits improved by approximately 1 log 10 to 0.3 RT‐PCRU per reaction when transcriptional enhancement and electrochemiluminescence (ECL) hybridization followed RT‐PCR. The method shows promise for improved detection sensitivity in instances where very low levels of virus contamination might be anticipated.