Embrapa Suínos E Aves, BR 153, Km 110, Concordia, SC, 89715-899, Brazil.
Embrapa Pecuária Sudeste, Rodovia Washington Luiz, Km 234, Fazenda Canchim, São Carlos, SP, 13560-970, Brazil.
Curr Microbiol. 2020 Jun;77(6):1043-1050. doi: 10.1007/s00284-020-01906-7. Epub 2020 Feb 4.
Infectious bursal disease (IBD) is an immunosuppressive viral disease of chickens, associated with severe economic losses and major threats to poultry production worldwide. Disease prevention programs rely on unequivocal identification of the pathogen, as well as vaccination programs. This study developed a sensitive, one-step, real-time, quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay using a hydrolysis probe system for infectious bursal disease virus (IBDV, VP1 gene) detection and quantification, which was compared to other routinely used diagnostic methods. The assay successfully detected IBD reference viruses and field isolates. The absence of cross-reactivity was detected with negative samples or with other avian viruses in the analytical specificity test. The detection limit of this assay was 70 RNA copies. RT-qPCR was more sensitive in the detection of serially diluted IBDV isolates compared to virus isolation. For clinical samples, the sensitivity and specificity values of RT-qPCR compared to enzyme-linked immunosorbent assay (ELISA) were 97.5% and 100%, respectively, and compared to histopathology, these values were 100% and 93.94%, respectively. RT-qPCR can provide a simple and reliable assay for IBDV surveillance programs and for evaluation of control strategies.
传染性囊病（IBD）是一种鸡的免疫抑制性病毒性疾病，与全球严重的经济损失和对家禽生产的重大威胁有关。疾病预防计划依赖于明确识别病原体以及疫苗接种计划。本研究开发了一种基于水解探针系统的敏感、一步、实时、定量逆转录聚合酶链反应（RT-qPCR）检测方法，用于传染性囊病病毒（IBDV，VP1 基因）的检测和定量，与其他常规使用的诊断方法进行了比较。该方法成功地检测到 IBD 参考病毒和田间分离株。在分析特异性试验中，未检测到与阴性样本或其他禽病毒的交叉反应性。该检测方法的检测限为 70 RNA 拷贝。与病毒分离相比，RT-qPCR 在检测连续稀释的 IBDV 分离株时更敏感。对于临床样本，与酶联免疫吸附试验（ELISA）相比，RT-qPCR 的敏感性和特异性值分别为 97.5%和 100%，与组织病理学相比，这些值分别为 100%和 93.94%。RT-qPCR 可为 IBDV 监测计划和控制策略的评估提供一种简单可靠的检测方法。
