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Journal of Clinical Microbiology, May 2009, p. 1517-1523, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.02011-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

PCR Diagnosis of Opisthorchis viverrini and Haplorchis taichui Infections in a Lao Community in an Area of Endemicity and Comparison of Diagnostic Methods for Parasitological Field Surveys{triangledown}

Leonore Lovis,1,2 Tippi K. Mak,2 Khampheng Phongluxa,2,3 Phonepasong Soukhathammavong,2,3 Somphou Sayasone,2,3 Kongsap Akkhavong,3 Peter Odermatt,2 Jennifer Keiser,4 and Ingrid Felger4*

Parasitology Laboratory, University Neuchâtel, Neuchâtel, Switzerland,1 Department of Public Health and Epidemiology, Swiss Tropical Institute, Basel, Switzerland,2 National Institute of Public Health, Vientiane, Lao People's Democratic Republic,3 Department of Medical Parasitology and Infection Biology, Swiss Tropical Institute, Basel, Switzerland4

Received 17 October 2008/ Returned for modification 26 December 2008/ Accepted 18 February 2009

Opisthorchiasis is a major public health problem in Southeast Asia. Affected individuals often have mixed infections with the liver fluke Opisthorchis viverrini and minute intestinal flukes such as Haplorchis taichui. The usual methods of diagnosing these infections involve the demonstration of fluke eggs in stool samples under light microscopy, but sensitivity and specificity are low. We developed two PCR tests that detect and discriminate between O. viverrini and H. taichui infections. PCR tests were validated by stool samples from purged individuals. We then applied the PCR tests to estimate the prevalence of O. viverrini and H. taichui infections from a random sample of individuals selected from a community in an area of endemicity in Khong District, Laos. PCR results were compared with those from the Kato-Katz (KK) method and the formalin-ether concentration technique (FECT). When validated with purge results, PCR tests of O. viverrini and H. taichui had sensitivities of 93.7% (95% confidence interval [CI], 85.8 to 97.9%) and 73.3% (95% CI, 60.3 to 83.9%) and could detect as little as 0.75 pg DNA and 1.32 ng DNA, respectively. The PCR-determined community prevalences of O. viverrini and H. taichui infections were 63.9% (95% CI, 54.1 to 72.9%) and 30.6% (95% CI, 22.1 to 40.2%), respectively. Using PCR as the gold standard to detect O. viverrini, three KK thick smears performed comparably well, whereas one KK smear and FECT were poorer (sensitivities of 91.4% [95% CI, 81.0 to 97.1%,], 62.3% [95% CI, 49.8 to 73.7%], and 49.3% [95% CI, 37.0 to 61.6%], respectively). PCR may be a valuable and sensitive diagnostic tool, particularly for low-intensity O. viverrini and H. taichui infections.

* Corresponding author. Mailing address: Department of Medical Parasitology and Infection Biology, Swiss Tropical Institute, Socinstrasse 57, 4002 Basel, Switzerland. Phone: 41 61 284 81 17. Fax: 41 61 271 86 54. E-mail: ingrid.felger{at}unibas.ch

{triangledown} Published ahead of print on 11 March 2009.

Journal of Clinical Microbiology, May 2009, p. 1517-1523, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.02011-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.



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