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外周血单个核细胞、血清及无细胞体腔积液的实时逆转录聚合酶链反应在猫传染性腹膜炎诊断中的比较

Comparison of real-time reverse transcriptase polymerase chain reaction of peripheral blood mononuclear cells, serum and cell-free body cavity effusion for the diagnosis of feline infectious peritonitis.

作者信息

Doenges Stephanie J, Weber Karin, Dorsch Roswitha, Fux Robert, Hartmann Katrin

机构信息

1 Clinic of Small Animal Medicine, LMU University of Munich, Germany.

2 Institute for Infectious Diseases and Zoonoses, LMU University of Munich, Germany.

出版信息

J Feline Med Surg. 2017 Apr;19(4):344-350. doi: 10.1177/1098612X15625354. Epub 2016 Jul 9.

Abstract

Objectives Diagnosis of feline infectious peritonitis (FIP) remains challenging, especially in cats without effusions. The objective of this study was to evaluate the sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction (RT-PCR) detecting feline coronavirus (FCoV) RNA in peripheral blood mononuclear cells (PBMCs) and serum in comparison with the same real-time RT-PCR in cell-free body cavity effusion. Methods This prospective case-control study included 92 cats. Forty-three cats had a definitive diagnosis of FIP, established either by histopathological examination (n = 28) or by positive immunofluorescence staining of FCoV antigen in macrophages of effusions (n = 11), or by both methods (n = 4). Forty-nine control cats had other diseases but similar clinical signs. Real-time RT-PCR was performed on PBMCs of 37 cats (21 cats with FIP, 16 controls), on serum of 51 cats (26 cats with FIP, 25 controls) and on cell-free body cavity effusion of 69 cats (36 cats with FIP, 33 controls). Sensitivity, specificity, positive and negative predictive value, including 95% confidence intervals (CI), were calculated. Results Real-time RT-PCR of PBMCs, serum and cell-free body cavity effusion showed a specificity of 100% (95% CI 79.4-100% in PBMCs, 86.3-100% in serum, 89.4-100% in cell-free body cavity effusion) and a sensitivity of 28.6% (95% CI 11.3-52.2%) in PBMCs, 15.4% (95% CI 4.4-34.9%) in serum and 88.9% (95% CI 73.9-96.9%) in cell-free body cavity effusion to diagnose FIP. Conclusions and relevance Although it is known that RT-PCR can often provide false-positive results in healthy cats, this real-time RT-PCR was shown to be a specific tool for the diagnosis of FIP when applied in a clinical setting. Sensitivity in cell-free body cavity effusion was high but low in PBMCs and serum. PBMC samples showed a higher sensitivity than serum samples, and are therefore a better choice if no effusion is present.

摘要

目的 猫传染性腹膜炎(FIP)的诊断仍然具有挑战性,尤其是在没有积液的猫中。本研究的目的是评估实时逆转录聚合酶链反应(RT-PCR)检测外周血单个核细胞(PBMC)和血清中猫冠状病毒(FCoV)RNA的敏感性和特异性,并与无细胞体腔积液中的相同实时RT-PCR进行比较。方法 这项前瞻性病例对照研究纳入了92只猫。43只猫被确诊为FIP,确诊方法包括组织病理学检查(n = 28)或积液巨噬细胞中FCoV抗原的免疫荧光染色阳性(n = 11),或两种方法均阳性(n = 4)。49只对照猫患有其他疾病但有相似的临床症状。对37只猫(21只FIP猫,16只对照)的PBMC、51只猫(26只FIP猫,25只对照)的血清以及69只猫(36只FIP猫,33只对照)的无细胞体腔积液进行实时RT-PCR检测。计算敏感性、特异性、阳性和阴性预测值,包括95%置信区间(CI)。结果 PBMC、血清和无细胞体腔积液的实时RT-PCR检测显示,其特异性为100%(PBMC中95%CI 79.4 - 100%,血清中86.3 - 100%,无细胞体腔积液中89.4 - 100%),PBMC诊断FIP的敏感性为28.6%(95%CI 11.3 - 52.2%),血清中为15.4%(95%CI 4.4 - 34.9%),无细胞体腔积液中为88.9%(95%CI 73.9 - 96.9%)。结论及相关性 虽然已知RT-PCR在健康猫中常可产生假阳性结果,但本实时RT-PCR在临床应用时被证明是诊断FIP的一种特异性工具。无细胞体腔积液中的敏感性高,而PBMC和血清中的敏感性低。PBMC样本的敏感性高于血清样本,因此在没有积液的情况下是更好的选择。

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