title: Sensitive and rapid detection of campylobacter species from stools of children with diarrhea in Japan by the loop-mediated isothermal amplification method.
abstract: We detected Campylobacter spp. in 5% (20/380) of diarrheal stool samples collected at an outpatient clinic in Kyoto using a commercial loop-mediated isothermal amplification (LAMP) kit with a fluorescent detection reagent after DNA extraction. The sensitivity and specificity were 100% in comparison with those of semi-nested PCR for the differentiation of Campylobacter jejuni and Campylobacter coli. Fourteen of the 20 samples were already determined as C. jejuni by the culture method. All 20 samples were also positive for C. jejuni by the PCR method. Among the 58 cultured samples, the sensitivity of the culture method against the LAMP method was 93.3% (14/15) and the specificity was 100% (43/43). The detection rate of Campylobacter spp. from the heated supernatants by the LAMP method was lower than that from the supernatant after DNA extraction. In total, 25% (5/20) of the Campylobacter-positive samples by the LAMP method were co-infected with norovirus (3/20), rotavirus (1/20), and human parechovirus (1/20), although no other bacterial co-infection was identified by the culture method. C. jejuni was mostly detected in children aged >5 years throughout the year. Based on these results, we concluded that care should be taken while diagnosing Campylobacter infection in children. Our newly modified LAMP method is a rapid, easy, and useful method for this diagnosis.
authors.affiliation: Division of Microbiology, Department of Pathology and Microbiology, Nihon University School of Medicine.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Ushijima
authors.forename: Shuichi
authors.initials: S
authors.lastname: Nishimura
authors.forename: Aksara
authors.initials: A
authors.lastname: Thongprachum
authors.forename: Yuko
authors.initials: Y
authors.lastname: Shimizu-Onda
authors.forename: Dinh Nguyen
authors.initials: DN
authors.lastname: Tran
authors.forename: Ngan Thi Kim
authors.initials: NT
authors.lastname: Pham
authors.forename: Sayaka
authors.initials: S
authors.lastname: Takanashi
authors.forename: Shuvra Kanti
authors.initials: SK
authors.lastname: Dey
authors.forename: Shoko
authors.initials: S
authors.lastname: Okitsu
authors.forename: Wataru
authors.initials: W
authors.lastname: Yamazaki
authors.forename: Masashi
authors.initials: M
authors.lastname: Mizuguchi
authors.forename: Satoshi
authors.initials: S
authors.lastname: Hayakawa
mesh_terms: D000293:Adolescent; D002167:Campylobacter / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D002169:Campylobacter Infections / Q000175:diagnosis* / Q000382:microbiology; D002648:Child; D002675:Child, Preschool; D060085:Coinfection / Q000175:diagnosis; D003967:Diarrhea / Q000175:diagnosis* / Q000382:microbiology; D005243:Feces / Q000382:microbiology*; D005260:Female; D006801:Humans; D007223:Infant; D007564:Japan; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D029322:Norovirus / Q000302:isolation & purification; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D030061:Parechovirus / Q000302:isolation & purification; D012401:Rotavirus / Q000302:isolation & purification; D012680:Sensitivity and Specificity