鸭圆环病毒和鹅圆环病毒三引物鉴别检测方法的建立

王梦鸽; 黄瑜; 傅秋玲; 程龙飞; 万春和; 刘荣昌; 陈红梅; 江南松; 梁齐章; 施少华; 林建生; 傅光华

doi:10.19303/j.issn.1008-0384.2023.01.002

鸭圆环病毒和鹅圆环病毒三引物鉴别检测方法的建立

Establishment of a Triple-primer Multiplex PCR Assay for Discriminative Detection of Duck and Goose Circoviruses

摘要

摘要:
目的建立一种可同时检测并鉴别鸭圆环病毒和鹅圆环病毒的PCR方法。
方法根据已发布的鸭圆环病毒和鹅圆环病毒基因组分子特征设计3条引物，应用3条引物建立了一种在一个反应管中可检测并鉴别鸭圆环病毒和鹅圆环病毒的三引物PCR方法。
结果该方法可特异性扩增鸭圆环病毒和鹅圆环病毒，并根据扩增片段大小鉴别病毒种类，检测最低限分别为110 、80 pg·μL⁻¹。应用该方法对临床疑似病例样品进行检测，其结果与用2种病毒对应的单病毒PCR检测方法一致。
结论建立的三引物PCR方法特异性强，敏感性高，丰富了水禽圆环病毒病的临床病例的诊断及流行病学监测的技术手段。

Abstract:
Objective A PCR assay was established for discriminative detection of duck circovirus (DuCV) and goose circovirus (GoCV).
Methods Three primers designed according to the genome sequences of DuCV and GoCV deposited in GenBank database were used for the development of a Triple-primer Multiplex PCR (tpm-PCR).
Results The established assay could specifically identify DuCV and GoCV by the amplified fragments within the detection limits of 110 pg·μL⁻¹ and 80 pg·μL⁻¹, respectively. The methodology was verified with the test results by PCR for the individual virus in clinical cases.
Conclusion The tpm-PCR assay was specific and sensitive as a discriminative detection method between DuCV and GoCV. It facilitated the clinical diagnosis and epidemiological monitoring on the circoviral cases in waterfowls.

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