Preparation of ELISA and Lateral Flow Kits for rapid Diagnosis of Mycoplasma gallisepticum in Poultry

Sherif Marouf,Rafik Sayed,Heidy Abo Elyazeed,Nayera M. Al-Atfeehy,Rasha Abotaleb

doi:10.1038/s41598-020-65848-7

Sherif Marouf, Rafik Sayed + Show 3 more

Open Access

https://doi.org/10.1038/s41598-020-65848-7

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Abstract

Avian mycoplasmas were mainly the cause of poultry industry economic losses; reduced meat and egg production and increases the antibiotic treatment cost. Mycoplasma gallisepticum (MG) infection is designated as infectious sinusitis of turkeys and chronic respiratory disease of chickens (gasping, depression, semi closed eyes, infraorbital sinuses edema and decrease in egg production). This study aimed to prepare, evaluate and Compare in-house ELISA kits and lateral flow assay (LFA) from a local strain of MG with commercial ELISA kits and PCR consequently. A total of 54 samples (27 tracheal swabs, 10 trachea and 17 lung) and 50 serum samples collected from birds suffering from chronic respiratory disease were tested by prepared in-house ELISA, commercial ELISA kits, PCR and LFA; a high correlation coefficient between in-house ELISA using whole antigen or sonicated antigen and commercial kit was recorded. Lateral Flow assay (LFA) performance indicate a low sensitivity (77.5%) but maintain a high specificity (92%) compared to PCR. The in-house ELISA kits and LFA prepared could be used as a fast diagnostic technique for detection of MG in Egypt. According to the available knowledge the prepared LFA for diagnosis of MG infection in chickens was developed for the first time in Egypt.

Highlights

Avian mycoplasmas are a major economic burden on the poultry industry; as decreased both feed conversion and quality of carcasses at slaughter
Different serological methods used for identification of avian mycoplasmas as ELISA and lateral flow assay
A high correlation coefficient (Pearson’s correlation coefficient (r) = 0.80, p = 0.009) depending on OD correlation values was detected between the prepared in-house ELISA whole antigen and Mycoplasma gallisepticum antibody test kit (ProFlock, Synbiotics Corporation, USA) as explained in Fig. (1) and Table (1)

Summary

Introduction

Avian mycoplasmas are a major economic burden on the poultry industry; as decreased both feed conversion and quality of carcasses at slaughter. It increase the mortality and loss of weight leading to severe economic losses. Serology and molecular methods are a diagnostic tools for diagnosis of avian mycoplasmas. Different serological methods used for identification of avian mycoplasmas as ELISA and lateral flow assay. ELISA plate coating antigen prepared from local isolate is cheap and produced reliable results to their commercial companion[4]. The following work aims to prepare and evaluate in-house ELISA kits and lateral flow assay (LFA) from a local strain of MG in comparison with commercial ELISA kits and PCR

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