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Development and evaluation of a colloidal gold immunochromatographic assay based on recombinant protein CatL1D for serodiagnosis of sheep fasciolosis

Published online by Cambridge University Press:  04 November 2019

W. Xifeng ,
Q. Mengfan ,
Z. Kai ,
Z. Guowu ,
L. Jing ,
W. Lixia ,
Q. Jun ,
M. Qingling ,
G. Shasha  and
H. Yunfu
...Show all authors
W. Xifeng
Affiliation:
College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832003, China
Q. Mengfan
Affiliation:
School of Biotechnology, Central South University, Changsha, Hunan 410012, China
Z. Kai
Affiliation:
College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832003, China
Z. Guowu
Affiliation:
College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832003, China
L. Jing
Affiliation:
College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832003, China
W. Lixia
Affiliation:
College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832003, China
Q. Jun
Affiliation:
College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832003, China
M. Qingling*
Affiliation:
College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832003, China
G. Shasha
Affiliation:
College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832003, China
H. Yunfu
Affiliation:
College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832003, China
C. Xuepeng
Affiliation:
State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu 730046, China
*
Author for correspondence: M. Qingling, E-mail: [email protected]
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Abstract

Fasciolosis is a zoonotic parasitic disease that seriously endangers the development of animal husbandry and human health. In order to develop a rapid serological diagnostic method for fasciolosis in ruminants, the CatL1D and CatB4 genes of Fasciola hepatica were amplified by reverse transcription polymerase chain reaction (PCR) and cloned, respectively, and then the CatL-B fusion gene (MeCatL-B) was constructed by gene splicing by overlap extension PCR technique. The recombinant rCatL1D, rCatB4 and rMeCatL-B proteins were then prepared by prokaryotic expression, respectively, and the recombinant protein with high specificity and sensitivity was screened via indirect enzyme-linked immunosorbent assay. Using the selected recombinant protein rCatL1D as a diagnostic antigen, we developed a colloidal gold immunochromatographic assay (CGIA) for detecting F. hepatica-specific antibodies, and 426 serum samples of slaughtered sheep were used to evaluate the sensitivity and specificity of F. hepatica CGIA assay. The results showed that the sensitivity and specificity of rCatL1D protein (100%, 96.67%) were higher than those of rCatB4 (94.29%, 80%) and rMeCatL-B (91.43%, 90%). Compared with the gold standard post-mortem inspection, the specificity and sensitivity of the CGIA method was 100% and 97%, respectively, and the consistency rate between these two methods was 99.3%. These results confirmed that the CGIA method based on rCatL1D protein could be a promising approach for rapid diagnosis of sheep fasciolosis because of its high sensitivity and specificity.

Keywords

Fasciola hepaticarCatL1DrCatB4rMeCatL-Bindirect ELISAreactogenicitycolloidal gold immunochromatography assay (CGIA)
Type
Research Paper
Information
Journal of Helminthology , Volume 94 , 2020 , e98
Copyright
Copyright © Cambridge University Press 2019 

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Footnotes

†

These authors contributed equally

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