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sELISA 检测泄殖腔拭子和病毒分离培养基中的 ALV p27。

Detection of ALV p27 in cloacal swabs and virus isolation medium by sELISA.

机构信息

Ministry of Education Key Lab for Avian Preventive Medicine, Yangzhou University, No. 12 East Wenhui Road, Yangzhou, Jiangsu, 225009, People's Republic of China.

Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, No. 12 East Wenhui Road, Yangzhou, Jiangsu, 225009, People's Republic of China.

出版信息

BMC Vet Res. 2019 Oct 30;15(1):383. doi: 10.1186/s12917-019-2150-z.

Abstract

BACKGROUND

Avian leukosis (AL), which is caused by avian leukosis virus (ALV), has led to substantial economic losses in the poultry industry. The kit used to detect all ALV-positive chickens in breeder flocks is very important for efficiently controlling AL. However, a new emerging ALV subtype is currently a severe challenge in the poultry industry.

RESULTS

In this paper, we compared different enzyme-linked immunosorbent assay (ELISA) kits for detecting p27 of ALV in the same batch of meconium samples. Different positive samples were further analyzed by PCR or virus isolation. The results showed that 36 positive samples among the 1812 chicken meconium samples could be detected by a sandwich ELISA (sELISA) kit, but only 17 positive samples could be identified by a commercial kit. To verify this result, cloacal swabs and viruses isolated from the positive chickens (2 days old) were used to detect the presence of p27. The results showed that the positive rate of p27 was 100% for the swabs and 40% for virus isolation. Surprisingly, PCR and sequence analysis revealed that the env gene of ALV in these positive samples belonged to the novel subgroup K (ALV-K).

CONCLUSION

These data not only demonstrate the relatively high sensitivity of the sELISA kit but also highlight the challenge of controlling ALV-K.

摘要

背景

由禽白血病病毒(ALV)引起的禽白血病(AL)给家禽业造成了巨大的经济损失。用于检测种鸡群中所有 ALV 阳性鸡的试剂盒对于有效控制 AL 非常重要。然而,目前新出现的 ALV 亚型是家禽业面临的严峻挑战。

结果

在本研究中,我们比较了同一批粪便样本中用于检测 ALV p27 的不同酶联免疫吸附测定(ELISA)试剂盒。对不同的阳性样本进一步通过 PCR 或病毒分离进行分析。结果表明,在 1812 份鸡粪便样本中,夹心 ELISA(sELISA)试剂盒可检测到 36 份阳性样本,但商业试剂盒仅能鉴定出 17 份阳性样本。为了验证这一结果,我们使用直肠拭子和从阳性鸡(2 日龄)中分离的病毒来检测 p27 的存在。结果表明,直肠拭子的 p27 阳性率为 100%,而病毒分离的阳性率为 40%。令人惊讶的是,PCR 和序列分析显示,这些阳性样本中 ALV 的 env 基因属于新型亚群 K(ALV-K)。

结论

这些数据不仅表明 sELISA 试剂盒具有相对较高的灵敏度,也突显了控制 ALV-K 的挑战。

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