title: Detection of Laribacter hongkongensis using species-specific duplex PCR assays targeting the 16S rRNA gene and the 16S-23S rRNA intergenic spacer region (ISR).
abstract: OBJECTIVE
  For the rapid detection of Laribacter hongkongensis, which is associated with human community-acquired gastroenteritis and traveller's diarrhoea, we developed a duplex species-specific PCR assay.
  
  
  RESULTS
  Full-length of the 16S-23S rRNA intergenic spacer region (ISR) sequences of 52 L. hongkongensis isolates were obtained by PCR-based sequencing. Two species-specific primer pairs targeting 16S rRNA gene and ISR were designed for duplex PCR detection of L. hongkongensis. The L. hongkongensis species-specific duplex PCR assay showed 100% specificity, and the minimum detectable level was 2.1 × 10(-2) ng μl(-1) genomic DNA which corresponds to 5000 CFU ml(-1).
  
  
  CONCLUSIONS
  The high specificity and sensitivity of the assay make it suitable for rapid detection of L. hongkongensis.
  
  
  CONCLUSIONS
  This species-specific duplex PCR method provides a rapid, simple, and reliable alternative to conventional methods to identify L. hongkongensis and may have applications in both clinical and environmental microbiology.
authors.affiliation: Hangzhou Center for Disease Control and Prevention, Jianqiao Town, Hangzhou, China.
authors.forename: L
authors.initials: L
authors.lastname: Shen
authors.forename: M
authors.initials: M
authors.lastname: Xiao
authors.forename: F
authors.initials: F
authors.lastname: Kong
authors.forename: M
authors.initials: M
authors.lastname: Brown
authors.forename: J
authors.initials: J
authors.lastname: Sun
authors.forename: Q
authors.initials: Q
authors.lastname: Kong
authors.forename: J
authors.initials: J
authors.lastname: Cha
authors.forename: H
authors.initials: H
authors.lastname: Xiang
authors.forename: H
authors.initials: H
authors.lastname: Xu
authors.forename: H
authors.initials: H
authors.lastname: Jin
authors.forename: L
authors.initials: L
authors.lastname: Wei
authors.forename: X
authors.initials: X
authors.lastname: Ni
mesh_terms: D000818:Animals; D001483:Base Sequence; D001717:Birds / Q000382:microbiology; D002347:Carps / Q000382:microbiology; D017714:Community-Acquired Infections / Q000382:microbiology; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D005759:Gastroenteritis / Q000382:microbiology; D006801:Humans; D008969:Molecular Sequence Data; D009346:Neisseriaceae / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D012338:RNA, Ribosomal, 23S / Q000235:genetics*; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D021903:DNA, Ribosomal Spacer; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S