Hostname: page-component-cd9895bd7-q99xh Total loading time: 0 Render date: 2024-12-26T20:40:38.427Z Has data issue: false hasContentIssue false
  • English
  • Français

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

Published online by Cambridge University Press:  01 May 2012

O. SANPOOL ,
P. M. INTAPAN ,
T. THANCHOMNANG ,
P. SRI-AROON ,
V. LULITANOND ,
L. SADAOW  and
W. MALEEWONG
O. SANPOOL
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
P. M. INTAPAN*
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
T. THANCHOMNANG
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Faculty of Medicine, Mahasarakham University, Mahasarakham 44000, Thailand
P. SRI-AROON
Affiliation:
Applied Malacology Center, Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand
V. LULITANOND
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
L. SADAOW
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
W. MALEEWONG
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Khon Kaen University, Khon Kaen 40002, Thailand Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
*
*Corresponding author: Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. Tel: +66 43 348387. Fax: +66 43202475. E-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

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.

Keywords

fecesratreal-time PCRSchistosoma mekongisnails
Type
Research Article
Information
Parasitology , Volume 139 , Issue 10 , September 2012 , pp. 1266 - 1272
Copyright
Copyright © Cambridge University Press 2012

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

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, 256–270. doi:10.1016/j.parint.2008.04.003.Google Scholar
Clerinx, J. and Van Gompel, A. (2011). Schistosomiasis in travellers and migrants. Travel Medicine and Infectious Disease 9, 6–24. doi: 10.1016/j.tmaid.2010.11.002.CrossRefGoogle ScholarPubMed
Galen, R. S. (1980). Predictive value and efficiency of laboratory testing. Pediatric Clinics of North America 27, 861–869.Google Scholar
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), 133–136. doi: 10.1590/S0074-02762006000900021.CrossRefGoogle ScholarPubMed
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.Google Scholar
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, 1–5. doi: 10.3347/kjp.2009.47.1.1.Google Scholar
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, 325–329. doi: 10.1016/j.exppara.2009.11.008.CrossRefGoogle ScholarPubMed
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, 25–33.Google ScholarPubMed
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, 1050–1055. doi: 10.4269/ajtmh.2009.09-0081.Google Scholar
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, 1123–1125.Google Scholar
Leshem, E., Meltzer, E., Marva, E. and Schwartz, E. (2009). Travel-related schistosomiasis acquired in Laos. Emerging Infectious Diseases 15, 1823–1826. doi: 10.3201/eid1511.090611.Google Scholar
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, 428–432.Google Scholar
Lyon, E. and Wittwer, C. T. (2009). LightCycler technology in molecular diagnostics. Journal of Molecular Diagnostics 11, 93–101. doi: 10.2353/jmoldx.2009.080094.CrossRefGoogle ScholarPubMed
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, 179–203. doi: 10.1016/S0065-308X(10)72007-8.CrossRefGoogle ScholarPubMed
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, 135–142. doi: 10.1016/j.parint.2004.01.004.CrossRefGoogle ScholarPubMed
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, 708–709. doi: 10.1645/0022-3395(2001)087[0708:TPAAHF]2.0.CO;2.Google Scholar
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, 509–513.CrossRefGoogle ScholarPubMed
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, 179–185. doi: 10.1016/j.trstmh.2007.10.011.Google Scholar
World Health Organization (2005). Report of the Scientific Working Group Meeting on schistosomiasis. TDR/SWG/07. World Health Organization, Geneva, Switzerland.Google Scholar