Discrimination between six species of canine microfilariae by a single polymerase chain reaction
Discrimination between six species of canine microfilariae by a single polymerase chain reaction
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DOI：
10.1016/j.vetpar.2005.10.013
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发表时间：
2006-02-18
影响因子：
2.6
通讯作者：
Alpizar, JLD
中科院分区：
文献类型：
--
作者：
Rishniw, M;Barr, SC;Alpizar, JLD
犬恶丝虫病通常通过特异性抗原检测和/或微丝蚴鉴定来诊断。然而，D.犬心丝虫和至少六种其他丝虫可产生犬微丝蚴病，犬心丝虫抗原检测呈阴性。为了解决两个诊断实验室对最近从欧洲进口到美国的抗原阴性微丝蚴血症犬的不一致诊断，我们开发了一种鉴定不同微丝蚴的简单分子方法，并随后验证了我们的方法对六个不同的丝虫已知感染狗通过扩增核糖体DNA间隔序列的聚合酶链反应使用常见的和物种-特异性引物，产物测序确认丝虫基因型。经鉴定，该虫为D.匍匐茎这是D.此外，我们检查了另外五只抗原阴性的微丝蚴血症犬的微丝蚴，并在每种情况下成功地鉴定了感染寄生虫。我们的诊断不同于最初的形态学诊断在这些情况下，证明形态学诊断的不准确性。在每种情况下，微丝蚴形态学鉴定为A。鉴定为D.通过分子生物学的方法来研究免疫系统。最后，我们证明，我们的PCR方法应扩增至少两个额外的丝虫（盘尾丝虫和曼森氏）的DNA，这表明这种方法可能是适合于家庭盘尾丝虫科的所有成员的基因分型。(c)2005 Elsevier B. V.保留所有权利。
Canine dirofilariasis caused by Dirofilaria immitis is usually diagnosed by specific antigen testing and/or identification of microfilariae. However, D. immitis and at least six other filariae can produce canine microfilaremias, with negative heartworm antigen tests. Discriminating these can be of clinical importance.To resolve discordant diagnoses by two diagnostic laboratories in an antigen-negative, microfilaremic dog recently imported into the US from Europe we developed a simple molecular method of identifying different microfilariae, and subsequently validated our method against six different filariae known to infect dogs by amplifying ribosomal DNA spacer sequences by polymerase chain reaction using common and species-specific primers, and sequencing the products to confirm the genotype of the filariae. We identified the filaria in this clog as D. repens. This is the first case of D. repens infection in the United States.Additionally, we examined microfilariae from five additional antigen-negative, microfilaremic dogs and successfully identified the infecting parasite in each case. Our diagnoses differed from the initial morphological diagnosis in three of these cases, demonstrating the inaccuracy of morphological diagnosis. In each case, microfilariae identified morphologically as A. reconditum were identified as D. immitis by molecular methods. Finally, we demonstrated that our PCR method should amplify DNA from at least two additional filariae (Onchocerca and Mansonella), suggesting that this method may be suitable for genotyping all members of the family Onchocercidae. (c) 2005 Elsevier B.V. All rights reserved.