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Evaluation of a SYBR Green Real-Time PCR Assay for Specific Detection of Pasteurella multocida in Culture and Tissue Samples from Sheep.

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Title Evaluation of a SYBR Green Real-Time PCR Assay for Specific Detection of Pasteurella multocida in Culture and Tissue Samples from Sheep.
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Creator Jyoti Kumar, GG Sonawane, Fateh Singh, S Jegaveera Pandian, Rajiv Kumar
 
Subject Pasteurella multocida, PCR, Real time PCR, Sensitivity, Specificity, Sheep, SYBR green. INTRODUCTION Respiratory
 
Description Not Available
Pasteurella multocida is one of the bacterial species involved in cases of ovine respiratory complex that has been implicated to cause
significant economic losses in sheep production system worldwide. The present study was undertaken with the aim of evaluating a
SYBR Green dye based real time PCR assay targeting KMT1 gene for the detection of P. multocida. The analytical specificity and
sensitivity of the PCR primers were evaluated. The test showed ten-fold more sensitivity than conventional PCR and detected down
to 275.5 fg/ μl of genomic DNA concentration, equivalent to 100 copies of KMT1 gene of P. multocida. The real-time PCR was found
to be specific for KMT1 gene of P. multocida, as no cross reactivity was detected with a variety of known bacterial isolates. A total of
52 ovine lung tissue samples were screened for P. multocida, which showed improved level of detection as compared to conventional
PCR. It is concluded that, this assay may be used as a valuable diagnostic tool for the rapid and specific detection of P. multocida.
By virtue of its high throughput format and its ability to accurately identify as well as quantify the bacterial DNA, the method may be
useful in large scale epidemiological studies and clarification of pathogenesis.
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Date 2021-07-15T09:51:46Z
2021-07-15T09:51:46Z
2020-08-01
 
Type Article
 
Identifier Not Available
Not Available
http://krishi.icar.gov.in/jspui/handle/123456789/48229
 
Language English
 
Relation Not Available;
 
Publisher Not Available