Masny Aleksander, Sałamatin Rusłan, Rozej-Bielicka Wioletta, Golab Elzbieta
Parasitol Res. 2016 Feb;115(2):511-25. doi: 10.1007/s00436-015-4767-6.
Dirofilaria repens is a parasite of animals and humans, transferred by mosquitoes. The assessment of the presence of D. repens-infected vertebrate hosts in the investigated area can be performed by xenomonitoring—detection of the parasite in blood-feeding arthropods. Our study aimed to evaluate PCR xenomonitoring of mosquitoes as a tool for dirofilariosis surveillance in Poland. We were also interested whether inter-study comparisons at the international level would be possible. Mosquitoes were collected in a single locality in Mazowsze province in Poland, in which between 12 and 20% of dogs were infected with D. repens and autochthonous human dirofilariosis was confirmed. All captured female mosquitoes were divided into pools; alternatively, single mosquitoes were analyzed; DNA was isolated and subjected to PCR and real-time PCR for detection of D. repens. The estimations of infection rates of mosquitoes with D. repens, based on PCR results, varied from 0 to 1.57% even between assays for detection of distinct fragments of the same marker—cytochrome oxidase subunit one gene. Polymorphisms of the DNA sequence within binding sites of the primers used in D. repens xenomonitoring assays, applied in European studies, were identified. Non-specific amplification of Setaria tundra (Nematoda: Onchocercidae) DNA occurred. Surveillance of dirofilariosis by PCR mosquito xenomonitoring is possible; however, the efficiency of the approach on territories where the prevalence of the disease among definitive hosts is lower than 12% remains unknown. Furthermore, mosquito infection rate estimations can be PCR assay dependent, which makes inter-study comparisons difficult. The results obtained in independent European xenomonitoring studies were contradictory. International collaboration would be required to establish a standardized set of assays for sensitive and specific xenomonitoring-based dirofilariosis surveillance.
匐行恶丝虫是一种人和动物共患的寄生虫，通过蚊子传播。在调查区域评估是否存在感染匐行恶丝虫的脊椎动物宿主，可以通过异体监测（即检测吸血节肢动物体内的寄生虫）来进行。我们的研究旨在评估通过PCR技术对蚊子进行异体监测，以此作为波兰恶丝虫病监测的一种工具。我们还想了解在国际层面上是否能够进行不同研究之间的比较。蚊子采自波兰马佐夫舍省的一个地点，该地区12%至20%的犬感染了匐行恶丝虫，且已确诊有本地感染的人体恶丝虫病病例。所有捕获的雌蚊被分成若干组；或者对单只蚊子进行分析；提取DNA后进行PCR和实时PCR检测，以确定是否存在匐行恶丝虫。基于PCR结果，对蚊子感染匐行恶丝虫的比率估计值在0至1.57%之间变化，即便在检测同一标记（细胞色素氧化酶亚基1基因）不同片段的检测之间也是如此。已确定欧洲研究中用于匐行恶丝虫异体监测检测的引物结合位点内DNA序列的多态性。出现了苔原腹腔丝虫（线虫纲：盘尾丝虫科）DNA的非特异性扩增。通过PCR技术对蚊子进行异体监测来开展恶丝虫病监测是可行的；然而，在终末宿主中该病患病率低于12%的地区，该方法的有效性仍不清楚。此外，蚊子感染率的估计可能取决于PCR检测方法，这使得不同研究之间难以进行比较。在欧洲独立开展的异体监测研究中得到的结果相互矛盾。需要开展国际合作，以建立一套标准化的检测方法，用于基于异体监测的敏感且特异的恶丝虫病监测。
