Journal of
Parasitology and Vector Biology

  • Abbreviation: J. Parasitol. Vector Biol.
  • Language: English
  • ISSN: 2141-2510
  • DOI: 10.5897/JPVB
  • Start Year: 2009
  • Published Articles: 204

Full Length Research Paper

Molecular xenomonitoring of trypanosomes in tsetse flies

Vincent P. Alibu*
  • Vincent P. Alibu*
  • Department of Biochemistry and Sports Sciences, Makerere University, P.O. Box 7062, Kampala, Uganda.
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John, C. K. Enyaru
  • John, C. K. Enyaru
  • Department of Biochemistry and Sports Sciences, Makerere University, P.O. Box 7062, Kampala, Uganda.
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Enock Matovu
  • Enock Matovu
  • College of Veterinary Medicine, Animal Resources and Biosecurity, Makerere University, P.O. Box 7062, Kampala, Uganda.
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Imna I. Malele
  • Imna I. Malele
  • Tsetse and Trypanosomiasis Research Institute (TTRI), Majani Mapana, Off Korogwe Road, P. O. Box 1026 Tanga, Tanzania.
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John E. Chisi
  • John E. Chisi
  • Tsetse and Trypanosomiasis Research Institute (TTRI), Majani Mapana, Off Korogwe Road, P. O. Box 1026 Tanga, Tanzania.
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Nicolas Mbongo
  • Nicolas Mbongo
  • Laboratoire National de Sante Publique, P.O. BOX 120 Brazzaville, Congo.
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Philemon Mansinsa
  • Philemon Mansinsa
  • National Program of Human African Trypanosomosis, P.O. BOX 7782 Kinshasa 1, Democratic Republic of Congo.
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El Rayah Intisar
  • El Rayah Intisar
  • Tropical Medicine Research Institute, P.O. BOX 1304, Khartoum, Sudan.
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Yassir Mohammed
  • Yassir Mohammed
  • Tropical Medicine Research Institute, P.O. BOX 1304, Khartoum, Sudan.
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Mubarak M. Abdelrahman
  • Mubarak M. Abdelrahman
  • Tropical Medicine Research Institute, P.O. BOX 1304, Khartoum, Sudan.
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Erneo B. Ochi
  • Erneo B. Ochi
  • Ministry of Animal Resources and Fisheries, P.O. Box 126 Juba, South Sudan.
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Yatta S. Lukaw
  • Yatta S. Lukaw
  • College of Natural Resources and Environmental Studies, University of Juba, P.O. Box 82 Juba, South Sudan.
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  •  Received: 02 February 2015
  •  Accepted: 15 May 2015
  •  Published: 31 July 2015

Abstract

Monitoring trypanosomes infections in wild-caught tsetse flies in a given area, is important in prediction of epidemic outbreaks and spread of disease, and could help focus control programs for areas requiring immediate attention in order to limit disease transmission and spread. The main objective of this study is to evaluate the recently developed RIME LAMP and PanTryp LAMP for screening large numbers of tsetse flies for trypanosomes and to assess their sensitivities and specificities for trypanosomes in endemic areas. Wild-caught tsetse flies were dissected and the mid-guts examined by microscopy. The mid-guts were pooled in fives (including one infected gut where applicable), homogenised and DNA extracted by Quiagen kits. TBR- and ITS-PCRs were carried out and examined under ethidium bromide-stained agarose gels while RIMELAMP and PanTryp LAMP were carried out and stained with SYBR green and also observed under ethidium bromide stained agarose gels. A total of 14912 tsetse flies identified as Glossina fuscipes fuscipes, Glossina. pallidipes, Glossina morsitans, Glossina. swynnertoni, Glossina fuscipes quazensis were trapped from the six different countries. Of these, 8789 were dissected. Both males and female tsetse flies had equal infection rates (12.2%) although overall infection rates varied with country. The highest number of infected tsetse flies was obtained by PanTryp LAMP followed by RIME LAMP, ITS-PCR, TBR-PCR and microscopy respectively. PanTryp LAMP was the most sensitive method followed by ITS-PCR, RIME LAMP and TBR-PCR respectively. However, ITS-PCR was the most specific followed by TBR-PCR, RIME LAMP and PanTryp LAMP respectively. Carrying out LAMP tests in the field provides the simplest and quickest means to estimate trypanosome infection rates in the vector tsetse flies. 

 
Key words: Xenomonitoring, trypanosome, tsetse fly, LAMP.