title: Development of a polymerase chain reaction applicable to rapid and sensitive detection of Clonorchis sinensis eggs in human stool samples.
abstract: Microscopic examination of eggs of parasitic helminths in stool samples has been the most widely used classical diagnostic method for infections, but tiny and low numbers of eggs in stool samples often hamper diagnosis of helminthic infections with classical microscopic examination. Moreover, it is also difficult to differentiate parasite eggs by the classical method, if they have similar morphological characteristics. In this study, we developed a rapid and sensitive polymerase chain reaction (PCR)-based molecular diagnostic method for detection of Clonorchis sinensis eggs in stool samples. Nine primers were designed based on the long-terminal repeat (LTR) of C. sinensis retrotransposon1 (CsRn1) gene, and seven PCR primer sets were paired. Polymerase chain reaction with each primer pair produced specific amplicons for C. sinensis, but not for other trematodes including Metagonimus yokogawai and Paragonimus westermani. Particularly, three primer sets were able to detect 10 C. sinensis eggs and were applicable to amplify specific amplicons from DNA samples purified from stool of C. sinensis-infected patients. This PCR method could be useful for diagnosis of C. sinensis infections in human stool samples with a high level of specificity and sensitivity.
authors.affiliation: Department of Parasitology and Inha Research Institute for Medical Sciences, Inha University School of Medicine, Incheon 400-712, Korea.
authors.forename: Pyo Yun
authors.initials: PY
authors.lastname: Cho
authors.forename: Byoung-Kuk
authors.initials: BK
authors.lastname: Na
authors.forename: Kyung Mi
authors.initials: KM
authors.lastname: Choi
authors.forename: Jin Su
authors.initials: JS
authors.lastname: Kim
authors.forename: Shin-Hyeong
authors.initials: SH
authors.lastname: Cho
authors.forename: Won-Ja
authors.initials: WJ
authors.lastname: Lee
authors.forename: Sung-Bin
authors.initials: SB
authors.lastname: Lim
authors.forename: Seok Ho
authors.initials: SH
authors.lastname: Cha
authors.forename: Yun-Kyu
authors.initials: YK
authors.lastname: Park
authors.forename: Jhang Ho
authors.initials: JH
authors.lastname: Pak
authors.forename: Hyeong-Woo
authors.initials: HW
authors.lastname: Lee
authors.forename: Sung-Jong
authors.initials: SJ
authors.lastname: Hong
authors.forename: Tong-Soo
authors.initials: TS
authors.lastname: Kim
mesh_terms: D000818:Animals; D003003:Clonorchiasis / Q000175:diagnosis* / Q000469:parasitology; D003004:Clonorchis sinensis / Q000235:genetics / Q000302:isolation & purification*; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000235:genetics; D005243:Feces / Q000469:parasitology*; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D010274:Parasitology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018626:Retroelements; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D017931:DNA Primers; D018509:DNA, Helminth; D018626:Retroelements
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