Skip to main content Accessibility help
×
Home

Development of a real-time PCR assay with fluorophore-labelled hybridization probes for detection of Schistosoma mekongi in infected snails and rat feces

  • O. SANPOOL (a1) (a2), P. M. INTAPAN (a1) (a2), T. THANCHOMNANG (a1) (a3), P. SRI-AROON (a4), V. LULITANOND (a1) (a5), L. SADAOW (a1) (a2) and W. MALEEWONG (a1) (a2)...

Summary

Schistosoma mekongi, a blood-dwelling fluke, is a water-borne parasite that is found in communities along the lower Mekong River basin, i.e. Cambodia and Lao People's Democratic Republic. This study developed a real-time PCR assay combined with melting-curve analysis to detect S. mekongi in laboratory setting conditions, in experimentally infected snails, and in fecal samples of infected rats. The procedure is based on melting-curve analysis of a hybrid between an amplicon from S. mekongi mitochondrion sequence, the 260 bp sequence specific to S. mekongi, and specific fluorophore-labelled probes. This method could detect as little as a single cercaria artificially introduced into a pool of 10 non-infected snails, a single cercaria in filtered paper, and 2 eggs inoculated in 100 mg of non-infected rat feces. All S. mekongi-infected snails and fecal samples from infected rats were positive. Non-infected snails, non-infected rat feces, and genomic DNA of other parasites were negative. The method gave high sensitivity and specificity, and could be applied as a fast and reliable tool for cercarial location in water environments in endemic areas and for epidemiological studies and eradication programmes for intermediate hosts.

Copyright

Corresponding author

*Corresponding author: Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. Tel: +66 43 348387. Fax: +66 43202475. E-mail: pewpan@kku.ac.th

References

Hide All
Attwood, S. W., Fatih, F. A., Campbell, I. and Upatham, E. S. (2008). The distribution of Mekong schistosomiasis, past and future: preliminary indications from an analysis of genetic variation in the intermediate host. Parasitology International 57, 256270. doi:10.1016/j.parint.2008.04.003.
Clerinx, J. and Van Gompel, A. (2011). Schistosomiasis in travellers and migrants. Travel Medicine and Infectious Disease 9, 624. doi: 10.1016/j.tmaid.2010.11.002.
Galen, R. S. (1980). Predictive value and efficiency of laboratory testing. Pediatric Clinics of North America 27, 861869.
Gomes, A. L., Melo, F. L., Werkhauser, R. P. and Abath, F. G. (2006). Development of a real time polymerase chain reaction for quantitation of Schistosoma mansoni DNA. Memórias do Instituto Oswaldo Cruz 101 (Suppl. 1), 133136. doi: 10.1590/S0074-02762006000900021.
Hung, Y. W. and Remais, J. (2008). Quantitative detection of Schistosoma japonicum cercariae in water by real-time PCR. PLoS Neglected Tropical Diseases 2, e337. doi: 10.1371/journal.pntd.0000337.
Ishikawa, H. and Ohmae, H. (2009). Modeling the dynamics and control of transmission of Schistosoma japonicum and S . mekongi in Southeast Asia. Korean Journal of Parasitology 47, 15. doi: 10.3347/kjp.2009.47.1.1.
Kato-Hayashi, N., Kirinoki, M., Iwamura, Y., Kanazawa, T., Kitikoon, V., Matsuda, H. and Chigusa, Y. (2010). Identification and differentiation of human schistosomes by polymerase chain reaction. Experimental Parasitology 124, 325329. doi: 10.1016/j.exppara.2009.11.008.
Kirinoki, M., Chigusa, Y., Ohmae, H., Sinuon, M., Socheat, D., Matsumoto, J., Kitikoon, V. and Matsuda, H. (2011). Efficacy of sodium metaperiodate (SMP)-ELISA for the serodiagnosis of schistosomiasis mekongi. Southeast Asian Journal of Tropical Medicine and Public Health 42, 2533.
Kjetland, E. F., Hove, R. J., Gomo, E., Midzi, N., Gwanzura, L., Mason, P., Friis, H., Verweij, J. J., Gundersen, S. G., Ndhlovu, P. D., Mduluza, T. and Van Lieshout, L. (2009). Schistosomiasis PCR in vaginal lavage as an indicator of genital Schistosoma haematobium infection in rural Zimbabwean women. American Journal of Tropical Medicine and Hygiene 81, 10501055. doi: 10.4269/ajtmh.2009.09-0081.
Le, T. H., Blair, D., Agatsuma, T., Humair, P. F., Campbell, N. J., Iwagami, M., Littlewood, D. T., Peacock, B., Johnston, D. A., Bartley, J., Rollinson, D., Herniou, E. A., Zarlenga, D. S. and McManus, D. P. (2000). Phylogenies inferred from mitochondrial gene orders-a cautionary tale from the parasitic flatworms. Molecular Biology and Evolution 17, 11231125.
Leshem, E., Meltzer, E., Marva, E. and Schwartz, E. (2009). Travel-related schistosomiasis acquired in Laos. Emerging Infectious Diseases 15, 18231826. doi: 10.3201/eid1511.090611.
Lier., T., Simonsen, G. S., Wang, T., Lu, D., Haukland, H. H., Vennervald, B. J., Hegstad, J. and Johansen, M. V. (2009). Real-time polymerase chain reaction for detection of low-intensity Schistosoma japonicum infections in China. American Journal of Tropical Medicine and Hygiene 81, 428432.
Lyon, E. and Wittwer, C. T. (2009). LightCycler technology in molecular diagnostics. Journal of Molecular Diagnostics 11, 93101. doi: 10.2353/jmoldx.2009.080094.
Muth, S., Sayasone, S., Odermatt-Biays, S., Phompida, S., Duong, S. and Odermatt, P. (2010). Schistosoma mekongi in Cambodia and Lao People's Democratic Republic. Advances in Parasitology 72, 179203. doi: 10.1016/S0065-308X(10)72007-8.
Ohmae, H., Sinuon, M., Kirinoki, M., Matsumoto, J. Chigusa, Y., Socheat, D. and Matsuda, H. (2004). Schistosomiasis mekongi: from discovery to control. Parasitology International 53, 135142. doi: 10.1016/j.parint.2004.01.004.
Strandgaard, H., Johansen, M. V., Pholsena, K., Teixayavong, K. and Christensen, N. O. (2001). The pig as a host for Schistosoma mekongi in Laos. Journal of Parasitology 87, 708709. doi: 10.1645/0022-3395(2001)087[0708:TPAAHF]2.0.CO;2.
Thanchomnang, T., Intapan, P. M., Lulitanond, V., Choochote, W., Manjai, A., Prasongdee, T. K. and Maleewong, W. (2008). Rapid detection of Brugia malayi in mosquito vectors using a real-time fluorescence resonance energy transfer PCR and melting curve analysis. American Journal of Tropical Medicine and Hygiene 78, 509513.
ten Hove, R. J., Verweij, J. J., Vereecken, K., Polman, K., Dieye, L. and van Lieshout, L. (2008). Multiplex real-time PCR for the detection and quantification of Schistosoma mansoni and S . haematobium infection in stool samples collected in northern Senegal. Transactions of the Royal Society of Tropical Medicine and Hygiene 102, 179185. doi: 10.1016/j.trstmh.2007.10.011.
World Health Organization (2005). Report of the Scientific Working Group Meeting on schistosomiasis. TDR/SWG/07. World Health Organization, Geneva, Switzerland.

Keywords

Development of a real-time PCR assay with fluorophore-labelled hybridization probes for detection of Schistosoma mekongi in infected snails and rat feces

  • O. SANPOOL (a1) (a2), P. M. INTAPAN (a1) (a2), T. THANCHOMNANG (a1) (a3), P. SRI-AROON (a4), V. LULITANOND (a1) (a5), L. SADAOW (a1) (a2) and W. MALEEWONG (a1) (a2)...

Metrics

Altmetric attention score

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed