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. 2014 Apr;6(2):115-21.
doi: 10.4103/0975-7406.126584.

A polymerase chain reaction and enzyme linked immunosorbent assay based approach for diagnosis and differentiation between vaccinated and infected cattle with Mycobacterium bovis

Mohamed Sabry  1 , Ahmed Elkerdasy  2
Affiliations

A polymerase chain reaction and enzyme linked immunosorbent assay based approach for diagnosis and differentiation between vaccinated and infected cattle with Mycobacterium bovis

Mohamed Sabry et al. J Pharm Bioallied Sci. 2014 Apr.

Abstract

Background: In most African and Arabic countries tuberculosis (TB) causes great economic losses in bovine species and constitutes serious zoonotic problem. As the traditional diagnostic method delay the research because of low sensitivity and specificity, a rapid method of diagnosis is of outmost importance.

Aim: The study was designed to evaluate the two rapid diagnostic methods of TB in cattle, further to differentiate between infected and bacillus Calmette-Guerin (BCG) vaccinated animals.

Materials and methods: Intradermal tuberculin test was applied to 300 cattle. Of these cattle, 15 cattle were vaccinated from cattle negative to tuberculin test with BCG. Blood samples were taken for lymphocyte separation to apply polymerase chain reaction (PCR) upon and for serum preparation for the enzyme-linked immunosorbent assay (ELISA) application, this blood collected from 65 cattle classified into three groups, viz. positive tuberculin test (35 animals), negative tuberculin test (15 animals), and vaccinated cow with BCG (15 animals). From blood samples lymphocytes were separated and the isolated lymphocytes were subjected to PCR and serum for ELISA application. Blood samples, specimens from lymph nodes and specific tissues were taken for PCR and for cultivation and isolation of Mycobacterium bovis.

Results and conclusions: The results of this study revealed that PCR can be used as rapid efficient and accurate diagnostic test in detection of ruminant TB. Moreover, cattle's ELISA reading showed higher sensitivity in positive tuberculin animals. However, the differentiations between vaccinated and infected animals not clear by using a single antigen only.

Keywords: Enzyme-linked immunosorbent assay; polymerase chain reaction; tuberculosis.

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Conflict of interest statement

Conflict of Interest: None declared.

Figures

Figure 1
Figure 1
Lanes 1-11 are positive, but lane 12 –ve control and lane 13 +ve control
Figure 2
Figure 2
Lanes 1-3, 6 and 8-11 are positive, but lane 12 –ve control and lane 13 +ve control
Figure 3
Figure 3
Lanes 2-5, 8-10 are positive, but lane 12 –ve control and lane 13 +ve control
Figure 4
Figure 4
Lanes 2-10 are positive, but lane 12 –ve control and lane 13 +ve control
Figure 5
Figure 5
Lanes from 5 to 10 are positive, but lane 12 –ve control and lane 13 +ve control
Figure 6
Figure 6
Lanes 2, 3 are positive, also lane 6-10 are positive, but lane 12 –ve control and lane 13 +ve control
See this image and copyright information in PMC

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