Background: Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. Methods: A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. Results: S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of the samples using pan-FMDV primers set, 1F/1R. Thirty-six samples were found positive and 7 negative by both the tests. The level of agreement between the tests was assessed by calculating the Kappa value, which was found to be fair (Kappa value = 0.303 and 95% CI = 0.089; 0.517) and significant (p = 0.009). However, 2 samples, which were found positive on S-ELISA tested negative on RT-PCR. This may be attributed to the presence of nucleotide mismatch(es) in the primer-binding sites that may have resulted in failure of amplification of the viral genome. The serotype-specific RT-PCR assays not only confirmed serotyping results of S-ELISA but were also able to establish serotype in 9 S-ELISA-negative but pan-FMDV RT-PCR-positive samples. Conclusions: The RT-PCR assay contributes significantly to establishing a quick, sensitive, and definitive diagnosis of FMD in resource-constrained countries. Samples giving negative results in S-ELISA should be tested in RT-PCR for the disease detection and virus typing.
Intervirology

中文翻译：

---

夹心酶联免疫吸附试验和逆转录聚合酶链反应在口蹄疫诊断中的评价

背景：口蹄疫（FMD）是偶蹄家畜和野生动物的一种传染性和高度传染性疾病，给畜牧业造成重大经济损失。对疾病进行快速可靠的诊断对于实施有效的控制措施至关重要。本研究比较了夹心酶联免疫吸附试验 (S-ELISA) 和常规逆转录聚合酶链反应 (RT-PCR) 对 FMD 的诊断。方法：使用 S-ELISA 和 RT-PCR 测定法检测了来自疑似 FMD 病例的总共 60 个上皮样本。通过计算 Kappa 值来评估测定之间的一致性水平。结果：S-ELISA 检测到 38 个（63%）样本为 FMD 病毒（FMDV）阳性。占主导地位的是，在 22 个 (57.9%) 的总样本中检测到血清型 O 呈阳性，而 9 个 (23.7%) 和 7 个 (18.4%) 样本分别被发现对血清型 A 和 Asia-1 呈阳性。RT-PCR 使用泛 FMDV 引物组 1F/1R 在 51 (85%) 个样本中检测到病毒基因组。两次测试均发现 36 个样本呈阳性，7 个样本呈阴性。通过计算 Kappa 值来评估测试之间的一致性水平，该值被认为是公平的（Kappa 值 = 0.303 和 95% CI = 0.089；0.517）和显着性（p= 0.009）。然而，在 S-ELISA 中发现阳性的 2 个样本在 RT-PCR 中检测为阴性。这可能归因于引物结合位点中核苷酸错配的存在，这可能导致病毒基因组扩增失败。血清型特异性 RT-PCR 检测不仅证实了 S-ELISA 的血清分型结果，而且还能够在 9 个 S-ELISA 阴性但泛 FMDV RT-PCR 阳性样品中建立血清型。结论： RT-PCR 检测有助于在资源有限的国家建立快速、灵敏和明确的 FMD 诊断。在 S-ELISA 中给出阴性结果的样品应在 RT-PCR 中检测疾病检测和病毒分型。
病毒学