Rapid Diagnosis of Bovine Brucellosis by Loop Mediated Isothermal Amplification (LAMP) assay

Abstract

Brucellosis is a zoonotic infection that cause abortions and infertility in livestock by effecting of especially genital organs like uterus, testes, mammary glands. In this study, it was aimed to use loop mediated isothermal amplification (LAMP), which is sensitive and specific molecular technique that based on amplification of BCSP31 gene, which is unique to organisms in the genus Brucella. In this study, a total of 57 materials including 20 milk samples and 37 aborted fetuses were tested by LAMP and classical bacterial culture for diagnosis of brucellosis. Thirteen (22.8%) out of 57 samples were found positive by LAMP while 10 samples (17.5%) yielded bacterial growth. As a conclusion, LAMP could be used satisfactorily for the diagnosis of brucellosis and the sensitivity can be increased by using two techniques together. We concluded that the bacterial culture is still gold standard for diagnosis of brucellosis although the sensitivity and specificity of the LAMP is high. 

Keywords

• Brucellosis,cattle,LAMP (Loop Mediated Isothermal Amplification),molecular diagnosis

Loop Mediated Isothermal Amplification (LAMP) Yöntemi ile Sığır Brusellozunun Hızlı Teşhisi

Abstract

Bruselloz, sığır, koyun, keçi, domuz ve koç gibi hayvanlarda, özellikle, testis, meme, uterus gibi genital organlara yerleşerek yavru atmalara ve infertiliteye neden olan zoonoz bir hastalıktır. Bu çalışmada bruselloz hastalığının çabuk tanısında BCSP31 genini temel alan duyarlı ve özgül özel bir moleküler teknik olan ilmek aracılı bir gen çoğaltma tekniğinin (loop mediated isothermal amplification), kullanılması amaçlandı. Bruselloz şüpheli hayvanlardan alınan 20 adet süt ve 37 adet aborte fötüs olmak üzere toplam 57 adet materyal LAMP ve kültür yöntemiyle incelendi. İncelenen materyallerin 13 adedi (%22,8) LAMP ile pozitif bulunurken, kültür yöntemi ile örneklerin 10’undan (%17,5) etken izole ve identifiye edildi. Sonuç olarak, LAMP tekniğinin brusellozun teşhisinde güven ile kullanılabileceği, LAMP testiyle beraber kültür yönteminin uygulanmasının teşhis sensitivitesinin artırılmasında yararlı olduğu saptandı. LAMP testinin spesifite ve sensitivitesinin yüksek olmasına rağmen hala kültür yönteminin altın standart olarak geçerliliğini sürdürdüğü düşünülmektedir.

Keywords

• Bruselloz,LAMP (Loop Mediated İsothermal Amplification),moleküler teşhis,sığır

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