Gunaydin  Elcin, Dakman  Asiye, Gulec  Metin, Cosar  Mustafa, Utuk  Armagan Erdem, Ozdemir  Umit, Laibinis  Victoria, 
    Multiplex-PCR for 2 Mycoplasmal Agents of Chicken Breeder Flocks and MG Vaccine Strain
Differentiation by mgc2-PCR RFLP,
    Research Journal of Poultry Sciences,
    Volume 13,Issue 3,
    2020,
    Pages 15-21,
    ISSN 1993-5285,
    rjpscience.2020.15.21,
    (https://makhillpublications.co/view-article.php?doi=rjpscience.2020.15.21)
    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
<i>Mycoplasma gallisepticum</i> (MG) or <i>Mycoplasma
synoviae</i> (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<sup>&#150;1</sup> and 10 CFU mL<sup>&#150;1</sup>,
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<sup>&#150;1</sup>, 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.
    Keywords: Mycoplasma gallisepticum;Mycoplasma synoviae;mPCR;mgc-2PCR RFLP