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巢式聚合酶链反应与血涂片检查在检测犬埃立克体和血小板无浆体中的比较

[Comparison of nested-PCR with blood smear examination in detection of Ehrlichia canis and Anaplasma platys in dogs].

作者信息

Ramos Carlos A N, Ramos Rafael A N, Araújo Flábio R, Guedes Daniel S, Souza Ingrid I F, Ono Tatiana M, Vieira Anahi S, Pimentel Danillo S, Rosas Eduardo O, Faustino Maria A G, Alves Leucio C

机构信息

Laboratório de Doenças Parasitárias, Departamento de Medicina Veterinária, Universidade Federal Rural de Pernambuco-UFRPE, Av. Dom Manoel de Medeiros s/n, Dois Irmãos, CEP 52171-900, Recife-PE, Brazil.

出版信息

Rev Bras Parasitol Vet. 2009 Dec;18 Suppl 1:58-62. doi: 10.4322/rbpv.018e1011.

Abstract

The clinical signs of Ehrlichia canis and Anaplasma platys infection are similar, and the diagnosis of these pathogens made by stained blood smears is poor due sensibility and specificity. On the other hand, the molecular diagnosis is highly sensitive and specific and nested-PCR have been optimized for accurate diagnosis these pathogens in dogs. At the veterinary teaching hospital, whole-blood samples with EDTA were obtained from 100 dogs and smears were made from blood samples for evaluation for intracellular parasites. For each sample, DNA was extracted and submitted to nPCR analysis for detection of E. canis and A. platys. The results of stained blood smears showed 9% of the animals were positive for E. canis and 21% for A. platys. Regarding of nPCR analysis, 57 and 55% of dogs were positive for E. canis and A. platys respectively. As compared to a nested PCR, the stained blood smears revealed false-negative results for both E. canis and A. platys. The results indicate that the nPCR is highly sensitive and specific for detection of both pathogens and the molecular diagnosis could be more useful at veterinary hospital.

摘要

犬埃立克体和血小板无浆体感染的临床症状相似,通过染色血涂片对这些病原体进行诊断时,由于敏感性和特异性较差,诊断效果不佳。另一方面,分子诊断具有高度的敏感性和特异性,巢式聚合酶链反应(nested-PCR)已得到优化,可用于准确诊断犬体内的这些病原体。在兽医教学医院,从100只犬采集了含乙二胺四乙酸(EDTA)的全血样本,并制作血涂片以评估细胞内寄生虫。对每个样本提取DNA,并进行巢式PCR分析以检测犬埃立克体和血小板无浆体。染色血涂片结果显示,9%的动物犬埃立克体呈阳性,21%的动物血小板无浆体呈阳性。关于巢式PCR分析,分别有57%和55%的犬犬埃立克体和血小板无浆体呈阳性。与巢式PCR相比,染色血涂片对犬埃立克体和血小板无浆体均显示出假阴性结果。结果表明,巢式PCR对两种病原体的检测具有高度的敏感性和特异性,分子诊断在兽医医院可能更有用。

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