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

Year 2018, Volume: 11 Issue: 1, 29 - 33, 30.06.2018

Osman Yaşar Tel Sevil Erdenliğ Gürbilek Oktay Keskin

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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