Development of real time PCR to study experimental mixed infections of T. congolense Savannah and T. b. brucei in Glossina morsitans morsitans

PloS One
Heba A AhmedKim Picozzi

Abstract

Tsetse flies are able to acquire mixed infections naturally or experimentally either simultaneously or sequentially. Traditionally, natural infection rates in tsetse flies are estimated by microscopic examination of different parts of the fly after dissection, together with the isolation of the parasite in vivo. However, until the advent of molecular techniques it was difficult to speciate trypanosomes infections and to quantify trypanosome numbers within tsetse flies. Although more expensive, qPCR allows the quantification of DNA and is less time consuming due to real time visualization and validation of the results. The current study evaluated the application of qPCR to quantify the infection load of tsetse flies with T. b. brucei and T. congolense savannah and to study the possibility of competition between the two species. The results revealed that the two qPCR reactions are of acceptable efficiency (99.1% and 95.6%, respectively), sensitivity and specificity and can be used for quantification of infection load with trypanosomes in experimentally infected Glossina morsitans morsitans. The mixed infection of laboratory Glossina species and quantification of the infection suggests the possibility that a form of competition ex...Continue Reading

References

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Jul 24, 2007·Experimental Parasitology·Kim PicozziSusan C Welburn
Mar 12, 2010·PLoS Pathogens·Lee R HainesMichael J Lehane

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Citations

Apr 14, 2017·BMC Veterinary Research·Reta D AbdiLuc Duchateau

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Methods Mentioned

BETA
PCR
dissection

Software Mentioned

GraphPad Prism
NanoDrop

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