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通过巢式PCR对全血及其组分作为诊断犬埃立克体病和无形体病的DNA来源进行评估。

An assessment of whole blood and fractions by nested PCR as a DNA source for diagnosing canine ehrlichiosis and anaplasmosis.

作者信息

Rotondano Tereza Emmanuelle de Farias, de Almeida Alzira Maria Paiva, Lustosa Elane Maria Camboim, Cordeiro Aline Antas, Camboim Expedito Kennedy Alves, de Azevedo Sérgio Santos, de Andrade Paulo Paes, de Melo Marcia Almeida

机构信息

Departamento de Ciências Biológicas, Universidade Federal de Pernambuco, Avenue Professor Moraes Rego, s/n, Cidade Universitária, 50.670-901 Recife, PE, Brazil.

出版信息

ScientificWorldJournal. 2012;2012:605743. doi: 10.1100/2012/605743. Epub 2012 Aug 22.

Abstract

Ehrlichiosis and anaplasmosis are tick-borne diseases. Ehrlichia canis and Anaplasma platys infect mainly white cells and platelets, respectively. The main DNA source for PCR is peripheral blood, but the potential of blood cell fractions has not been extensively investigated. This study aims at assessment of whole blood (WB) and blood fractions potential in nested PCR (nPCR) to diagnose canine ehrlichiosis and anaplasmosis. The 16S rRNA gene was amplified in 71.4, 17.8, 31.57, and 30% of the WB, granulocyte (G), mononuclear cells (M), and buffy coat (BC) samples. Compared to the WB, the sensitivity of the PCR was 42.86% for the M, and BC fractions, 21.43% for the G, and 33.33% for the blood clot (C). There was fair agreement between the WB and M, BC and C, and slight with the G. Fair agreement occurred between the nPCR and morulae in the blood smear. One animal was coinfected with A. platys and E. canis. This study provided the first evidence of A. platys infection in dogs in Paraíba, Brazil, and demonstrated that WB is a better DNA source than blood fractions to detect Ehrlichia and Anaplasma by nPCR, probably because of the plasma bacterial concentration following host cell lysis.

摘要

埃立克体病和无形体病是蜱传播疾病。犬埃立克体和血小板无形体分别主要感染白细胞和血小板。聚合酶链反应(PCR)的主要DNA来源是外周血,但血细胞成分的潜力尚未得到广泛研究。本研究旨在评估全血(WB)和血液成分在巢式PCR(nPCR)中诊断犬埃立克体病和无形体病的潜力。在71.4%的WB样本、17.8%的粒细胞(G)样本、31.57%的单核细胞(M)样本和30%的血沉棕黄层(BC)样本中扩增出16S rRNA基因。与WB相比,PCR对M和BC成分的敏感性为42.86%,对G的敏感性为21.43%,对血凝块(C)的敏感性为33.33%。WB与M、BC与C之间有较好的一致性,与G之间一致性稍差。nPCR与血涂片上的桑葚体之间有较好的一致性。1只动物同时感染了血小板无形体和犬埃立克体。本研究提供了巴西帕拉伊巴州犬感染血小板无形体的首个证据,并表明通过nPCR检测埃立克体和无形体时,WB作为DNA来源比血液成分更好,这可能是由于宿主细胞裂解后血浆中的细菌浓度所致。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/1546/3432355/dc532e7ab6c7/TSWJ2012-605743.001.jpg

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