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Background <p>Mycoplasma synoviae is a serious pathogen of poultry in worldwide causing severe economic losses. Phase-variable haemagglutinin, a highly expressed lipoprotein, serves as a key membrane immunogen of M. synoviae. The objective of this research was to develop a chimeric multiepitope protein of M. synoviae for potential application in ELISA test.</p> Methods <p>Specific linear B-cell epitopes from the vlhA and MS2/28 proteins were carefully chosen. The multiepitope protein was produced in BL21-DE3 strain of E.coli and purified using an affinity purification method.</p> Results <p>A substantial amount of recombinant proteins was successfully expressed in a soluble form and purified, yielding 71mg protein per 1 liter bacterial culture. An indirect rMEVlhA-MS2/28-ELISA method was developed for detecting M. synoviae antibodies. 163 M. synoviae positive, negative and vaccinated chicken serum samples from the Veterinary Organization, veterinary diagnostic laboratories, and poultry disease department of RAZI institute were obtained and were used to evaluation of performance of rMEVlhA-MS2/28-ELISA.</p> Conclusions <p>Our study highlights the promise of utilizing rMEVlhA-MS2/28 protein, offering novel insights for the development of M. synoviae indirect ELISA test. The strong specificity of rMEVlhA-MS2/28 protein may be attributed to the selection of specific epitopes and removing the G-related Albumin-binding module in the vlhA protein. The rMEVlhA-MS2/28-ELISA serves as a convenient, sensitive, and highly specific diagnostic tool that can aid in rapid diagnosis and epidemiological investigations. This chimeric protein shows promise in detecting specific antibodies against M. synoviae, potentially contributing to improved detection methods.</p>

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Designing and Production of Novel rMEVlhA-MS2/28 Multiepitopes Protein Applicable in Indirect ELISA

  • 期刊论文

  • 发表日期：

    2025年9月10日

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  • Majid Esmaelizad,
  • Maryam Javadnejad,
  • Marzieh Ghollasi,
  • Mohsen Bashashati,
  • Mohammad Ranjbar,
  • Rasool Madani,
  • Fatemeh Gholami,
  • Aysan Parvizi,
  • Nazanin Esmaelizad,
  • Ali Mollaie

摘要

Background

Mycoplasma synoviae is a serious pathogen of poultry in worldwide causing severe economic losses. Phase-variable haemagglutinin, a highly expressed lipoprotein, serves as a key membrane immunogen of M. synoviae. The objective of this research was to develop a chimeric multiepitope protein of M. synoviae for potential application in ELISA test.

Methods

Specific linear B-cell epitopes from the vlhA and MS2/28 proteins were carefully chosen. The multiepitope protein was produced in BL21-DE3 strain of E.coli and purified using an affinity purification method.

Results

A substantial amount of recombinant proteins was successfully expressed in a soluble form and purified, yielding 71mg protein per 1 liter bacterial culture. An indirect rMEVlhA-MS2/28-ELISA method was developed for detecting M. synoviae antibodies. 163 M. synoviae positive, negative and vaccinated chicken serum samples from the Veterinary Organization, veterinary diagnostic laboratories, and poultry disease department of RAZI institute were obtained and were used to evaluation of performance of rMEVlhA-MS2/28-ELISA.

Conclusions

Our study highlights the promise of utilizing rMEVlhA-MS2/28 protein, offering novel insights for the development of M. synoviae indirect ELISA test. The strong specificity of rMEVlhA-MS2/28 protein may be attributed to the selection of specific epitopes and removing the G-related Albumin-binding module in the vlhA protein. The rMEVlhA-MS2/28-ELISA serves as a convenient, sensitive, and highly specific diagnostic tool that can aid in rapid diagnosis and epidemiological investigations. This chimeric protein shows promise in detecting specific antibodies against M. synoviae, potentially contributing to improved detection methods.