Volume 5, Issue 18 (4-2015)                   NCMBJ 2015, 5(18): 49-54 | Back to browse issues page

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Mohammadpour M. Isolation and identification of Mycoplasma agalactiae by culture and polymerase chain reaction (PCR) from milk of sheep in Masal, Guilan. NCMBJ. 2015; 5 (18) :49-54
URL: http://ncmbjpiau.ir/article-1-626-en.html
, Mahdi_mohamadpour@yahoo.com
Abstract:   (7662 Views)
Aim and Background: Contagious agalactiae (C.A) is a serious disease syndrome of sheep and goat characterized by mastitis and subsequent failure of milk production, arthritis, abortion and kerato-conjunctivitis. Mycoplasma agalactiae (M. agalactiae) is the main cause of the disease in sheep and goat. The aim of this study was isolation and identification of M. agalactiae with culture and polymerase chain reaction (PCR) assay from milk of sheep in Masal, Iran
Material and Methods: A total of 25 samples were collected from milk secretion of sheep. All samples were cultured in PPLO broth supplemented for M. agalactiae isolation. The bacteria DNAs were extracted by High Pure PCR Template Preparation kit and the PCR assay was applied for the detection of Mycoplasma genus in 270bp fragment in 16S rRNA gene and M. agalactiae in 375bp fragment in lipoprotein gene from culture as well as clinical samples.
Results: Of the 25 samples, 9 (36%) yielded one of the potentially pathogenic Mycoplasmas evaluated for using PPLO agar culture diagnostic method, and 17 (68%) yielded one of the potentially pathogenic Mycoplasmas evaluated for using Mycoplasma genus PCR as diagnostic method, and 5 (29/4%) of the samples yielded M. agalactiae by using M. agalactiae PCR as diagnostic method. Of the 25 samples, 8 samples were culture positive and PCR positive, 7 samples were culture negative and PCR negative whereas 9 samples were culture negative and PCR positive and only one sample was culture positive and PCR negative.
Conclusion: The result showed that PCR can be used as an alternative method instead of culturing to identify M. agalactiae. M. agalactiae was detected for the first time from milk of sheep in Masal, Iran and it was not the main factor of C.A there.
Full-Text [PDF 540 kb]   (2093 Downloads)    
Type of Study: Research Article | Subject: Cellular and molecular
Received: 2015/05/31 | Accepted: 2015/05/31 | Published: 2015/05/31

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