羊痘病毒抗体胶体金免疫层析试纸条的研制与初步应用

doi:10.11843/j.issn.0366-6964.2025.07.029

Abstract

Abstract:

This study aimed to establish a rapid, simple, specific and sensitive colloidal gold-based universal detection method for antibodies against Capripoxvirus. By constructing a recombinant vector and expressing and purifying the recombinant 122 protein of the Capripoxvirus in a prokaryotic system, rabbit polyclonal antibodies were prepared. The 122 protein was conjugated with colloidal gold as the gold-labeled antigen. Then, the 122 protein and rabbit polyclonal antibodies were coated onto a nitrocellulose membrane (NC) as the test line (T line) and control line (C line), respectively. After optimizing the conditions, a colloidal gold immunochromatographic test strip for detecting Capripoxvirus antibodies was developed. The results showed that the prokaryotically expressed recombinant 122 protein was approximately 32 ku in size. The developed colloidal gold test strip could specifically detect Capripoxvirus antibodies within 12 minutes; there was no cross-reaction with positive sera of other common livestock diseases. The sensitivity for detecting sheeppox virus lumpy skin disease virus and goat-pox virus positive sera was 1 ∶64, 1 ∶128, and 1 ∶128, respectively, which was comparable to the sensitivity of the commercially available indirect ELISA diagnostic kit for Capripoxvirus antibodies (titers 1 ∶128, 1 ∶256, 1 ∶256). The detection of 130 clinical serum samples was compared with the results of the commercial kit, and the kappa value was 0.93, indicating a high degree of consistency. This study successfully developed a universal colloidal gold immunochromatographic test strip for detecting Capripoxvirus antibodies, which has high sensitivity and specificity, as well as low cost, rapid detection, simple operation, and easy interpretation of results, providing practical application value for on-site detection of Capripoxvirus.

Key words: Capripoxvirus, protein 122, polyclonal antibodies, colloidal gold test strips

CLC Number:

S852.659.1

HE Yindi, SHI Zhengwang, SHI Xintai, CHEN Jie, LIAO Huancheng, ZHANG Fan, LUO Juncong, ZHU Yuqian, XI Tao, LI Shuaipeng, WANG Chuan, TIAN Hong, ZHENG Haixue. Development and Preliminary Application of Colloidal Gold Immunochromatographic Test Strips for Antibodies against Capripoxvirus[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(7): 3368-3377.

Figures/Tables 7

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References 30

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储存时间/月Storage time(month)	室温(18~25 ℃)Room temperature(18~25 ℃)	4 ℃环境4 ℃ environment
0	+	+
1	+	+
3	+	+
6	+	+
9	-	+
12	-	+
15	-	-

方法Method		Biovetest ELISA试剂盒Biovetest ELISA kit
方法Method		阳性Positive	阴性Negative	合计Total
试纸条Test strip	阳性Positive	41	0	41
	阴性Negative	4	85	89
	合计Total	45	85	130
Kappa值Kappa value	0.93
Kappa value

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Development and Preliminary Application of Colloidal Gold Immunochromatographic Test Strips for Antibodies against Capripoxvirus

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