Development of real time PCR to study experimental mixed infections of T. congolense Savannah and T. b. brucei in Glossina morsitans morsitans.
Development of real time PCR to study experimental mixed infections of T. congolense Savannah and T. b. brucei in Glossina morsitans morsitans.
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DOI：
10.1371/journal.pone.0117147
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Picozzi K
中科院分区：
文献类型：
--
作者：
Ahmed HA;MacLeod ET;Welburn SC;Picozzi K
采采蝇能够自然地或实验地同时或顺序地获得混合感染。传统上，采采蝇的自然感染率是通过解剖后对蝇的不同部位进行显微镜检查以及体内寄生虫的分离来估计的。然而，直到分子技术的出现，它是很难物种锥虫感染和量化锥虫的数量在采采蝇。虽然更昂贵，但qPCR允许DNA的定量，并且由于结果的真实的时间可视化和验证而耗时更少。本研究评价了qPCR在定量采采蝇T. B. brucei和T. congolense萨凡纳，并研究这两个物种之间竞争的可能性。结果表明，两种qPCR反应具有可接受的效率（分别为99.1%和95.6%）、灵敏度和特异性，并且可用于定量实验感染的桑舌虫中锥虫的感染负荷。实验室Glossina种的混合感染和感染的定量表明T. B. brucei和T.刚果大萨凡纳，我们用它们共存于苍蝇的中肠。
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 exists between the isolates of T. b. brucei and T. congolense savannah that we used when they co-exist in the fly midgut.