Development of a Novel PmpD-N ELISA for Chlamydia psittaci Infection

LIU Shan Shan; CHU Jun; ZHANG Qiang; SUN Wei; ZHANG Tian Yuan; HE Cheng

doi:10.3967/bes2016.041

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Volume 29 Issue 5

May 2016

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Article Navigation > Biomedical and Environmental Sciences > 2016 > 29(5): 315-322

LIU Shan Shan, CHU Jun, ZHANG Qiang, SUN Wei, ZHANG Tian Yuan, HE Cheng. Development of a Novel PmpD-N ELISA for Chlamydia psittaci Infection[J]. Biomedical and Environmental Sciences, 2016, 29(5): 315-322. doi: 10.3967/bes2016.041

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LIU Shan Shan, CHU Jun, ZHANG Qiang, SUN Wei, ZHANG Tian Yuan, HE Cheng. Development of a Novel PmpD-N ELISA for Chlamydia psittaci Infection[J]. Biomedical and Environmental Sciences, 2016, 29(5): 315-322. doi: 10.3967/bes2016.041

Development of a Novel PmpD-N ELISA for Chlamydia psittaci Infection

doi: 10.3967/bes2016.041

Key Lab of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China
Tongren Polytechnic College, Tongren 554300, Guizhou, China

Funds: the National Natural Science Foundation of China under Grant No.31272542%Ministry of Science and Technology (MoST) grant 2012AA101302

Abstract

ObjectiveChlamydia psittaci is an avian respiratory pathogen and zoonotic agent.The wide prevalence ofC. psittaci poses a threat to the poultry industry and its employees. However, few commercial kits are available for detecting avian antibodies excluding the in-house ELISA kit. In this study, we developed a novel ELISA kit for detecting antibodies againstC. psittaci based on the N-terminal fragment of polymorphic outer membrane protein D (PmpD-N) as the coating antigen.
Methods The antigen concentrations, primary antibody, and cut-off value were determined and optimized. The ELISA, designated PmpD-N ELISA, was assessed for sensitivity, specificity, and concordance using sera samples from 48 experimentally infected and 168 uninfected SPF chickens.
Results The sensitivity and specificity of PmpD-N ELISA were 97.9%, 100%, respectively, while the concordance was 98.1% as compared to that of MOMP-ELISA. No cross-reaction with positive sera for other avian pathogens was found. Using PmpD-N ELISA, 799/836 clinical samples were positive, including 93.0% and 98.1% positivity in layers and broilers, respectively.
Conclusion These data indicate that indirect ELISA with PmpD-N as the antigen candidate is a promising approach for the surveillance ofC. psittaci infection.
- Chlamydia psittaci,
- Indirect ELISA,
- N-terminal fragment of polymorphic membrane protein D,
- Seroprevalence
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通讯作者: 陈斌, bchen63@163.com

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沈阳化工大学材料科学与工程学院沈阳 110142

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Development of a Novel PmpD-N ELISA for Chlamydia psittaci Infection

doi: 10.3967/bes2016.041

Key Lab of Animal Epidemiology and Zoonosis of Ministry of Agriculture, College of Veterinary Medicine, China Agricultural University, Beijing 100193, China

Tongren Polytechnic College, Tongren 554300, Guizhou, China

Funds: the National Natural Science Foundation of China under Grant No.31272542%Ministry of Science and Technology (MoST) grant 2012AA101302

Key words:

Chlamydia psittaci /
Indirect ELISA /
N-terminal fragment of polymorphic membrane protein D /
Seroprevalence

Abstract: ObjectiveChlamydia psittaci is an avian respiratory pathogen and zoonotic agent.The wide prevalence ofC. psittaci poses a threat to the poultry industry and its employees. However, few commercial kits are available for detecting avian antibodies excluding the in-house ELISA kit. In this study, we developed a novel ELISA kit for detecting antibodies againstC. psittaci based on the N-terminal fragment of polymorphic outer membrane protein D (PmpD-N) as the coating antigen.
Methods The antigen concentrations, primary antibody, and cut-off value were determined and optimized. The ELISA, designated PmpD-N ELISA, was assessed for sensitivity, specificity, and concordance using sera samples from 48 experimentally infected and 168 uninfected SPF chickens.
Results The sensitivity and specificity of PmpD-N ELISA were 97.9%, 100%, respectively, while the concordance was 98.1% as compared to that of MOMP-ELISA. No cross-reaction with positive sera for other avian pathogens was found. Using PmpD-N ELISA, 799/836 clinical samples were positive, including 93.0% and 98.1% positivity in layers and broilers, respectively.
Conclusion These data indicate that indirect ELISA with PmpD-N as the antigen candidate is a promising approach for the surveillance ofC. psittaci infection.

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