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01.12.2015 | Research article | Ausgabe 1/2016 Open Access

BMC Infectious Diseases 1/2016

Characterization of Sarcoptes scabiei cofilin gene and assessment of recombinant cofilin protein as an antigen in indirect-ELISA for diagnosis

Zeitschrift:
BMC Infectious Diseases > Ausgabe 1/2016
Autoren:
Yu Zheng, Ran He, Manli He, Xiaobin Gu, Tao Wang, Weimin Lai, Xuerong Peng, Guangyou Yang
Wichtige Hinweise

Competing interests

The authors have declared that no competing interests exist.

Authors’ contributions

YZ participated in the design of the study, manuscript writing and performed the statistical analysis; RH participated in the write of the manuscript, RH and MH participated in the collection of mites samples; RH and YZ participated in experiment and discussion; YZ participated in sequence alignment, XG and TW helped to draft the manuscript; WL and XP provided the support of experiment animals and serum samples; GY participated in the design of study and have given final approval of the version; All authors read and approved the final manuscript.

Abstract

Background

Scabies impairs the health of humans and animals and causes heavy economic losses. Traditional diagnostic methods for scabies are inefficient and ineffective, and so far there is no commercial immunodiagnostic or molecular based test for scabies.

Methods

Here, we used recombinant Sarcoptes scabiei cofilin protein as an antigen to establish indirect ELISA. S. scabiei cofilin is highly homologous to Dermatophagoides farinae Der f 31 allergen (90 % identity). The S. scabiei cofilin gene was cloned and expressed in Escherichia coli to obtain recombinant protein. Western blotting and fluorescence immunohistochemistry were carried out, and we established an indirect ELISA method and detected 33 serum samples from scabies infected rabbits and 30 serum samples from naïve rabbits.

Results

Western blotting demonstrated that S. scabiei cofilin possessed good immunogenicity and fluorescence immunohistochemistry showed the S. scabiei cofilin is widespread in the splanchnic area of mites. In ELISA, a cut-off value of 0.188 was determined to judge experimental positive and negative serum values. Specificity and sensitivity of the ELISA were 87.9 and 83.33 %, respectively.

Conclusions

Recombinant S. scabiei cofilin showed potential value as a diagnostic antigen. The ELISA method established could be used in clinical diagnosis and provide experimental information in minimal or asymptomatic infection.
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