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Research Journal of Poultry Sciences

ISSN: Online
ISSN: Print 1993-5285
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Multiplex-PCR for 2 Mycoplasmal Agents of Chicken Breeder Flocks and MG Vaccine Strain Differentiation by mgc2-PCR RFLP

Gunaydin Elcin, Dakman Asiye, Gulec Metin, Cosar Mustafa, Utuk Armagan Erdem, Ozdemir Umit and Laibinis Victoria
Page: 15-21 | Received 21 Sep 2022, Published online: 21 Sep 2022

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https://doi.org/10.36478/rjpscience.2020.15.21

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Abstract

Due to the economic impact of Mycoplasma infection in poultry, it is essential to have a fast, reliable and accurate diagnostic test to diagnose the infection. Multiplex-PCR (mPCR) is advantageous in that a single swab can be used to identify the presence of either Mycoplasma gallisepticum (MG) or Mycoplasma synoviae (MS), testing can be completed in half the time, using fewer materials resulting in lower expense. The objectives of this study are two-fold: to optimize a mPCR for the detection of MG and MS from a single tracheal swab in order to investigate the presence of MG and MS in breeder flocks in Turkey and to differentiate the MG vaccine strains, ts-11 and 6/85 from field infection. Sensitivity of the mPCR was determined to be 6 colony forming units (CFU) mL–1 and 10 CFU mL–1, respectively, from pure MG S6 and MS WVU1853 cultures. In artificially spiked samples with pure MG S6 and MS WVU1853 cultures, sensitivity decreased to 60 and 100 CFU mL–1, respectively. A total of 900 tracheal swab samples were collected from nine chicken breeder flocks, three flocks each from Ankara, Bolu and Eskisehir provinces. Swabs were pooled into groups of 5, (180 pools) and were examined for the presence of MG and MS by mPCR and bacteriology. Testing revealed, 1/180 (0.55%) was found MS positive by both mPCR and culture. While 6/180 (3.88%) were determined MS positive, solely by mPCR. Differentiation of 6/85 and ts11 MG vaccine strains from field strains was achieved by mgc2 PCR-RFLP using HaeII restriction endonuclease enzyme.

How to cite this article:

Gunaydin Elcin, Dakman Asiye, Gulec Metin, Cosar Mustafa, Utuk Armagan Erdem, Ozdemir Umit and Laibinis Victoria. Multiplex-PCR for 2 Mycoplasmal Agents of Chicken Breeder Flocks and MG Vaccine Strain Differentiation by mgc2-PCR RFLP.
DOI: https://doi.org/10.36478/rjpscience.2020.15.21
URL: https://www.makhillpublications.co/view-article/1993-5285/rjpscience.2020.15.21