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Sensitive and rapid detection of campylobacter species from stools of children with diarrhea in Japan by the loop-mediated isothermal amplification method

Authors :
Sayaka Takanashi
Satoshi Hayakawa
Shoko Okitsu
Yuko Shimizu-Onda
Masashi Mizuguchi
Hiroshi Ushijima
Shuvra Kanti Dey
Shuichi Nishimura
Wataru Yamazaki
Dinh Nguyen Tran
Ngan Thi Kim Pham
Aksara Thongprachum
Source :
Japanese journal of infectious diseases. 67(5)
Publication Year :
2014

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.

Details

ISSN :
18842836
Volume :
67
Issue :
5
Database :
OpenAIRE
Journal :
Japanese journal of infectious diseases
Accession number :
edsair.doi.dedup.....cc45b853c28d1084992f663aac7f663c