Rapid Molecular Detection of Opisthorchis viverrini in Human Fecal Samples by Real-Time Polymerase Chain Reaction

Pewpan M. Intapan Faculty of Medicine and Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand; Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand

Search for other papers by Pewpan M. Intapan in
Current site
Google Scholar
PubMed
Close
,
Tongjit Thanchomnang Faculty of Medicine and Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand; Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand

Search for other papers by Tongjit Thanchomnang in
Current site
Google Scholar
PubMed
Close
,
Viraphong Lulitanond Faculty of Medicine and Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand; Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand

Search for other papers by Viraphong Lulitanond in
Current site
Google Scholar
PubMed
Close
,
Phunthira Pongsaskulchoti Faculty of Medicine and Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand; Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand

Search for other papers by Phunthira Pongsaskulchoti in
Current site
Google Scholar
PubMed
Close
, and
Wanchai Maleewong Faculty of Medicine and Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen, Thailand; Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand

Search for other papers by Wanchai Maleewong in
Current site
Google Scholar
PubMed
Close
Restricted access

Real-time fluorescence resonance energy transfer (FRET) polymerase chain reaction (PCR) supplemented with melting curve analysis is a highly sensitive and fast method offering a high throughput. We report the development of a real-time FRET PCR for molecular detection of Opisthorchis viverrini in human fecal samples. The diagnostic sensitivity, specificity, accuracy, and positive and negative predictive values of this method were 97.5%, 100%, 98.9%, 100%, and 98.2%, respectively. The sensitivity was not significantly different from that of the quantified formalin-ethyl acetate concentration technique, the gold standard (P > 0.05). The procedure has potential for diagnosis of human opisthorchiasis in disease-endemic areas, for large epidemiologic investigations involving at risk populations, and monitoring eradication programs of the liver fluke, which causes hepatobiliary diseases and induces cholangiocarcinoma.

Author Notes

  • 1

    World Health Organization, 1995. Control of foodborne trematode infections. Report of a WHO Study Group. World Health Organ Tech Rep Ser 849 :1–57.

  • 2

    Chai JY, Darwin Murrell K, Lymbery AJ, 2005. Fish-borne parasitic zoonoses: status and issues. Int J Parasitol 35 :1233–1254.

  • 3

    Yossepowitch O, Gotesman T, Assous M, Marva E, Zimlichman R, Dan M, 2004. Opisthorchiasis from imported raw fish. Emerg Infect Dis 10 :2122–2126.

  • 4

    Woolf A, Green J, Levine JA, Estevez EG, Weatherly N, Rosenberg E, Frothingham T, 1984. A clinical study of Laotian refugees infected with Clonorchis sinensis or Opisthorchis viverrini. Am J Trop Med Hyg 33 :1279–1280.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 5

    Schwartz DA, 1986. Cholangiocarcinoma associated with liver fluke infection: a preventable source of morbidity in Asian immigrants. Am J Gastroenterol 81 :76–79.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 6

    Peng HW, Chao HL, Fan PC, 1993. Imported Opisthorchis viverrini and parasite infections from Thai labourers in Taiwan. J Helminthol 67 :102–106.

  • 7

    Mairiang E, Haswell-Elkins MR, Mairiang P, Sithithaworn P, Elkins DB, 1993. Reversal of biliary tract abnormalities associated with Opisthorchis viverrini infection following praziquantel treatment. Trans R Soc Trop Med Hyg 87 :194–197.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 8

    Schwartz DA, 1980. Helminths in the induction of cancer: Opisthorchis viverrini, Clonorchis sinensis and cholangiocarcinoma. Trop Geogr Med 32 :95–100.

  • 9

    Harinasuta T, Riganti M, Bunnang D, 1984. Opisthorchis viverrini infection: pathogenesis and clinical features. Arzneimittelforschung 34 :1167–1169.

  • 10

    Pungpak S, Chalermrut K, Harinasuta T, Viravan C, Schelp PF, Hempfling A, Schlattmann P, Bunnag D, 1994. Opisthorchis viverrini infection in Thailand: symptoms and signs of infection-a population-based study. Trans R Soc Trop Med Hyg 88 :561–564.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 11

    Sripa B, Kaewkes S, Sithithaworn P, Mairiang E, Laha T, Smout M, Pairojkul C, Bhudhisawasdi V, Tesana S, Thinkamrop B, Bethony JM, Loukas A, Brindley PJ, 2007. Liver fluke induces cholangiocarcinoma. PLoS Med 4 :1148–1154.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 12

    Wongratanacheewin S, Pumidonming W, Sermswan RW, Maleewong W, 2001. Development of a PCR-based method for the detection of Opisthorchis viverrini in experimentally infected hamsters. Parasitology 122 :175–180.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 13

    Wongratanacheewin S, Pumidonming W, Sermswan RW, Pipitgool V, Maleewong W, 2002. Detection of Opisthorchis viverrini in human stool specimens by PCR. J Clin Microbiol 40 :3879–3880.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 14

    Stensvold CR, Saijuntha W, Sithithaworn P, Wongratanacheewin S, Strandgaard H, Ornbjerg N, Johansen MV, 2006. Evaluation of PCR based coprodiagnosis of human opisthorchiasis. Acta Trop 97 :26–30.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 15

    Duenngai K, Sithithaworn P, Rudrappa UK, Iddya K, Laha T, Stensvold CR, Strandgaard H, Johansen MV, 2008. Improvement of PCR for detection of Opisthorchis viverrini DNA in human stool samples. J Clin Microbiol 46 :366–368.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 16

    Umesha KR, Kumar S, Parvathi A, Duenngai K, Sithithaworn P, Karunasagar I, Karunasagar I, 2008. Opisthorchis viverrini: detection by polymerase chain reaction (PCR) in human stool samples. Exp Parasitol 120 :353–356.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 17

    Intapan PM, Thanchomnang T, Lulitanond V, Pongsaskulchoti P, Maleewong W, 2008. Detection of Opisthorchis viverrini in infected bithynid snails by real-time fluorescence resonance energy transfer PCR-based method and melting curve analysis. Parasitol Res 103 :649–655.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 18

    Intapan PM, Thanchomnang T, Lulitanond V, Phongsaskulchoti P, Maleewong W, 2008. Real-time fluorescence resonance energy transfer PCR with melting curve analysis for the detection of Opisthorchis viverrini in fish intermediate hosts. Vet Parasitol 157 :65–71.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 19

    Suksumek N, Leelawat K, Leelawat S, Russell B, Lek-Uthai U, 2008. TaqMan real-time PCR assay for specific detection of Opisthorchis viverrini DNA in Thai patients with hepatocellular carcinoma and cholangiocarcinoma. Exp Parasitol 119 :217–224.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 20

    Lyon E, Wittwer CT, 2009. LightCycler technology in molecular diagnostics. J Mol Diagn 11 :93–101.

  • 21

    Elkins DB, Haswell-Elkins M, Anderson RM, 1986. The epidemiology and control of intestinal helminths in the Pulicat Lake region of southern India. I. Study design and pre- and post-treatment observations on Ascaris lumbricoides infection. Trans R Soc Trop Med Hyg 80 :774–792.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 22

    Sermswan R, Mongkolsuk S, Panyim S, Sirisinha S, 1991. Isolation and characterization of Opisthorchis viverrini specific DNA probe. Mol Cell Probes 5 :399–407.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 23

    Thanchomnang T, Intapan PM, Lulitanond V, Choochote W, Manjai A, Prasongdee TK, Maleewong W, 2008. Rapid detection of Brugia malayi in mosquito vectors using a real-time fluorescence resonance energy transfer PCR and melting curve analysis. Am J Trop Med Hyg 78 :509–513.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 24

    Intapan PM, Maleewong W, 2006. Opisthorchis viverrini: influence of maternal infection in hamsters on offspring infected with homologous parasite and their IgG antibody response. Exp Parasitol 113 :67–74.

    • PubMed
    • Search Google Scholar
    • Export Citation
  • 25

    Galen RS, 1980. Predictive value and efficiency of laboratory testing. Pediatr Clin North Am 27 :861–869.

  • 26

    Sirisinha S, Chawengkirttikul R, Haswell-Elkins MR, Elkins DB, Kaewkes S, Sithithaworn P, 1995. Evaluation of a monoclonal antibody-based enzyme linked immunosorbent assay for the diagnosis of Opisthorchis viverrini infection in an endemic area. Am J Trop Med Hyg 52 :521–524.

    • PubMed
    • Search Google Scholar
    • Export Citation
Past two years Past Year Past 30 Days
Abstract Views 355 327 27
Full Text Views 273 2 0
PDF Downloads 62 2 0
 
 
 
 
Affiliate Membership Banner
 
 
Research for Health Information Banner
 
 
CLOCKSS
 
 
 
Society Publishers Coalition Banner
Save