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比色原位杂交与定量实时聚合酶链反应在福尔马林固定石蜡包埋犬皮肤样本中诊断[病原体名称未给出]感染的比较研究

Comparative Study of Colorimetric In Situ Hybridization and Quantitative Real-Time Polymerase Chain Reaction for Diagnosis of Infection by in Dogs in Formalin-Fixed and Paraffin-Embedded Skin.

作者信息

Silva Ricardo Gonçalves, Kiupel Matti, Langohr Ingeborg Maria, Wise Annabel, Pereira Sandro Antonio, Gonçalves Natália Pedra, da Conceição Greice Maria Silva, Ferreira Luiz Cláudio, de Campos Monique Paiva, Miranda Luciana de Freitas Campos, Figueiredo Fabiano Borges, de Oliveira Raquel de Vasconcellos Carvalhaes, Keidel Lucas, Menezes Rodrigo Caldas

机构信息

Laboratory of Clinical Research on Dermatozoonoses in Domestic Animals, Evandro Chagas National Institute of Infectious Diseases, Oswaldo Cruz Foundation, Av. Brasil, 4365, Rio de Janeiro 21040-360, Brazil.

Veterinary Diagnostic Laboratory, Michigan State University, 4125 Beaumont Road, Lansing, MI 48910, USA.

出版信息

Trop Med Infect Dis. 2024 Apr 22;9(4):91. doi: 10.3390/tropicalmed9040091.

Abstract

The zoonotic visceral leishmaniasis is caused by the protozoan and dogs are reservoirs for this parasite. For the diagnosis of at the species level in dogs in formalin-fixed, paraffin-embedded skin (FFPES) samples, colorimetric in situ hybridization (CISH) and quantitative real-time polymerase chain reaction (qPCR) are options, but their sensitivities are not well established. Therefore, the aim of this study was to determine the sensitivity of these two techniques in FFPES for the diagnosis of the infection in dogs using culture as the reference standard. The FFPES of 48 dogs with cutaneous infection by confirmed by culture and by multilocus enzyme electrophoresis were examined by CISH and qPCR using specific probes for . The sensitivities of qPCR, CISH and their combination were, respectively, 77.0%, 58.0% and 83.3%. The sensitivities of qPCR in dogs with and without clinical signs were, respectively, 74.2% and 82.4%. The sensitivities of CISH in dogs with and without clinical signs were, respectively, 61.3% and 52.9%. The CISH and qPCR showed satisfactory sensitivities for the diagnosis of in the FFPES of dogs, even in dogs without clinical signs, and their combination increases the sensitivity for this diagnosis.

摘要

人畜共患内脏利什曼病由原生动物引起,狗是这种寄生虫的宿主。对于福尔马林固定、石蜡包埋皮肤(FFPES)样本中犬类的该物种水平诊断,比色原位杂交(CISH)和定量实时聚合酶链反应(qPCR)是可选方法,但它们的敏感性尚未得到充分证实。因此,本研究的目的是使用培养作为参考标准,确定这两种技术在FFPES中对犬类 感染诊断的敏感性。使用针对 的特异性探针,通过CISH和qPCR对48只经培养和多位点酶电泳确诊为皮肤感染 的犬的FFPES进行检测。qPCR、CISH及其联合检测的敏感性分别为77.0%、58.0%和83.3%。有临床症状和无临床症状犬的qPCR敏感性分别为74.2%和82.4%。有临床症状和无临床症状犬的CISH敏感性分别为61.3%和52.9%。CISH和qPCR对犬FFPES中 的诊断显示出令人满意的敏感性,即使在无临床症状的犬中也是如此,并且它们的联合使用提高了该诊断的敏感性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fcf0/11054412/e389fbab86a0/tropicalmed-09-00091-g001.jpg

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