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| Content Provider | Springer Nature Link |
|---|---|
| Author | Prusty, Bikash R. Chaudhuri, Pallab Chaturvedi, V. K. Saini, Mohini Mishra, B. P. Gupta, Praveen K. |
| Copyright Year | 2016 |
| Abstract | Several pathogens including Brucella spp. are shed in semen of infected bulls and can be transmitted to cows through contaminated semen during artificial insemination. The present study reports omp2a and bcsp31 gene based loop-mediated isothermal amplification (LAMP) assays for detection of Brucella genomic DNA in semen from infected bulls. The positive results could be interpreted visually by change in colour of reaction mixture containing hydroxyl naphthol blue (HNB) dye from violet to sky blue. LAMP assays based on omp2a and bcsp31 could detect as little as 10 and 100 fg of B. abortus S19 genomic DNA, respectively. Sensitivity of omp2a and bcsp31 LAMP assays for direct detection of organisms in bovine semen was 2.28 × 10$^{1}$ CFU and 2.28 × 10$^{2}$ CFU of B. abortus S19 in spiked bovine semen, respectively. The omp2a LAMP assay was found equally sensitive to TaqMan probe based real-time PCR and 100 times more sensitive than conventional PCR in identifying Brucella in spiked semen. The diagnostic applicability of the omp2a LAMP assay was evaluated with seventy-nine bovine semen samples and results were re-evaluated through TaqMan probe based real-time PCR and conventional PCR. Taken together, the omp2a LAMP assay is easy to perform, rapid and sensitive in diagnosis of Brucella spp. in bovine semen. |
| Starting Page | 142 |
| Ending Page | 147 |
| Page Count | 6 |
| File Format | |
| ISSN | 00468991 |
| Journal | Indian Journal of Microbiology |
| Volume Number | 56 |
| Issue Number | 2 |
| e-ISSN | 09737715 |
| Language | English |
| Publisher | Springer India |
| Publisher Date | 2016-01-05 |
| Publisher Institution | Association of Microbiologists of India |
| Publisher Place | New Delhi |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Loop-mediated isothermal amplification assay Bovine brucellosis Bovine semen omp2a bcsp31 Microbiology Medical Microbiology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Microbiology |
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