title: Genogroup-specific PCR primers for detection of Norwalk-like viruses.
abstract: Norwalk-like viruses (NLV) are a major causative agent of nonbacterial gastroenteritis. There are still many NLV strains that are refractory to gene amplification by ordinary reverse transcription-polymerase chain reaction. This is due mainly to the genetic diversity among NLV, especially mismatches in the primer sequences, which limits this technique in clinical utility. In this study, improved primer sets based on the capsid region, to detect both genogroup I and II NLV by genogroup-specific manner, were developed. When stool specimens from gastroenteritis patients, that were positive for NLV by electron microscopy, were tested by this new primer set, all specimens were positive by RT-PCR. Primers described previously for RdRp and capsid protein were capable of amplifying the specimens by 31 and 77%, respectively. Therefore, new primer sets are extremely useful for the amplification and rapid diagnosis of nonbacterial gastroenteritis due to NLV as well as for epidemiological studies.
authors.affiliation: Section of Infectious Disease, R&D Center, BML, 1361-1, Matoba, Kawagoe, Saitama 350-1101, Japan.
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Kojima
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Kageyama
authors.forename: Shuetsu
authors.initials: S
authors.lastname: Fukushi
authors.forename: Fuminori B
authors.initials: FB
authors.lastname: Hoshino
authors.forename: Michiyo
authors.initials: M
authors.lastname: Shinohara
authors.forename: Kazue
authors.initials: K
authors.lastname: Uchida
authors.forename: Katsuro
authors.initials: K
authors.lastname: Natori
authors.forename: Naokazu
authors.initials: N
authors.lastname: Takeda
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Katayama
mesh_terms: D001483:Base Sequence; D017250:Caliciviridae Infections / Q000453:epidemiology / Q000821:virology*; D002213:Capsid / Q000235:genetics*; D036022:Capsid Proteins; D017931:DNA Primers; D004279:DNA, Viral; D004196:Disease Outbreaks; D005759:Gastroenteritis / Q000453:epidemiology / Q000821:virology*; D006801:Humans; D007564:Japan / Q000453:epidemiology; D008969:Molecular Sequence Data; D029322:Norovirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods; D012367:RNA, Viral / Q000032:analysis*; D012324:RNA-Dependent RNA Polymerase / Q000235:genetics*; D012680:Sensitivity and Specificity
chemical_list: D036022:Capsid Proteins; D017931:DNA Primers; D004279:DNA, Viral; C121877:Norwalk virus capsid; D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase