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An improved method for the detection of Norwalk‐like caliciviruses in environmental samples
Author(s) -
Loisy F.,
Le Cann P.,
Pommepuy M.,
Le Guyader F.
Publication year - 2000
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1365-2672.2000.00837.x
Subject(s) - norwalk virus , caliciviridae , biology , microbiology and biotechnology , virology , rna , gel electrophoresis , oligonucleotide , real time polymerase chain reaction , norovirus , virus , chemistry , gene , genetics
An original magnetic beads RNA capture was developed for the detection of Norwalk‐like virus by RT‐PCR. The same oligonucleotide was used both for capture and reverse transcription of the viral RNA. The optimization studies showed that the most important parameter for sensitivity is the biotin‐binding capacity of the beads. This method was found to be efficient for eliminating inhibitors in sewage samples compared with the classic RT‐PCR. Moreover, the sensitivity was greatly enhanced, allowing the detection of 42% positive sample after gel electrophoresis, which is fourfold greater than classic RT‐PCR (11%). Beads‐RT‐PCR sensitivity is the same as classic RT‐PCR and hybridization. Thus, this method, which is easy to perform, should be of particular interest for developing quantitative RT‐PCR and sequencing.