Diagnosis and detection of Mycoplasma Synoviae from commercial poultry flocks using polymeras chain reaction (PCR) based on amplification of VlhA gene

Subject Areas : Journal of Comparative Pathobiology

معصومه مقامی ¹ , سیدعلی پوربخش ² , علیرضا همایونی مهر ³ , حمیدرضا مهاجرانی ⁴ , عباس اشتری ⁵ , محمدعلی بیات زاده ⁶

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Received: 2012-09-26 Accepted : 2012-12-28 Published : 2013-02-19

Keywords: Mycoplasma Synoviae, Variable Lipoprotein Heamagglu, VlhA Gene, PCR, Isolation, Diagnosis, detection,

Abstract :

Mycoplasma Synoviae is a major avian pathogen and cases many economic losses on poultryindustry in Iran, Mycoplasma Synoviae synthesizes Variable lipoprotein heamagglutinin(VlhA),is believed to play a major in pathogenesis of the disease. The objective of the present study wasto diagnosis and detection of Mycoplasma Synoviae isolates based on the vlhA gene.This studywas designed to detect MS through culture isolation and polymerase chain reaction (PCR) assayto demonstrated the involvement of MS infection in trachea and the lung/air sac samples takenfrom commercial poultry flocks of provinces: Tehran, Qazvin and Markazi with clinical signsof the disease. Moreover the specific primers to the Mycoplasma genuse from the primescomplementary to the single-copy conserved 5´ end of vlhA gene were used for detection of MSby vlhA-PCR. PCR results, in addition to identification of Mycoplasma specie, revealedvariable sizes of 350-400bp for each isolates. Of the 43 swabs 28(65%) yielded one of thepotentially pathogenic Mycoplasmas evaluated for using PPLO agar culture diagnostic method,and 33(76%) yielded one of the potentially pathogenic Mycoplasmas evaluated for Mycoplasmagenuse PCR as diagnostic metod and 24(55%) swabs yielded Mycoplasma Synoviae for usingvlhA PCR as diagnostic metod. The results of this study demonstrated that PCR based onamplification of conserved 5́ end of vlhA gene is an efficient tool for detection of MycoplasmaSynoviae isolates, as well as due to contain of RI, RII and RIII regions its can with top sensitiveand operative, be used for to diagnosis and detection of Mycoplasma Synoviae isolates.