Sensitive detection of human Caliciviridae by RT-PCR.
A semi-nested reverse transcriptase-polymerase chain reaction (RT-PCR) was developed for the detection of human Caliciviridae. The method was evaluated on faecal samples from patients with gastroenteritis sent to the Norwegian National Institute of Public Health for routine diagnosis by direct electron microscopy (EM). Of 166 samples, 49 were found to contain Caliciviridae by EM, while 7 samples contained other viruses. A total of 74 samples was positive by PCR, including all the samples with EM detectable Caliciviridae, while specimens containing other agents were negative. Phylogenetic analysis of RNA sequences from 14 Norwegian samples indicated that the viruses present in Norway are evenly distributed when compared to sequences of human Caliciviridae from other countries. The PCR primers should therefore be useful for samples from other regions. The phylogenetic analysis did not cluster viruses with a calici-like morphology, but mingled them with sequences from Norwalk-like viruses, indicating that the two morphological types do not represent separate genogroups.
Stene-Johansen
K
K
Department of Virology, National Institute of Public Health, Oslo, Norway.
Grinde
B
B
D001483:Base Sequence; D002139:Caliciviridae / Q000145:classification / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D017250:Caliciviridae Infections / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis*; D005243:Feces / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D004279:DNA, Viral; D012324:RNA-Dependent RNA Polymerase