Methods for precise molecular detection of probiotic microflora : using adjusted molecular biology protocols, primer sets and PCR assays.
Lactobacillus sp. is probiotic bacteria for which many detection methods were envisaged. However, culture-based methods failed to achieve specific detection of this bacterium due to its presence in mixed bacterial complex communities. The PCR assay was optimized to detect and quantify Lactobacillus...
| Главные авторы: | , , , |
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| Формат: | Статья |
| Язык: | English English |
| Опубликовано: |
Asian Network for Scientific Information
2010
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| Online-ссылка: | http://psasir.upm.edu.my/id/eprint/13937/1/Methods%20for%20precise%20molecular%20detection%20of%20probiotic%20microflora.pdf |
| _version_ | 1825945332283867136 |
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| author | Abu Bakar, Fatimah Abdulamir, Ahmed Sahib Nordin, Noordiana Yoke, T. S. |
| author_facet | Abu Bakar, Fatimah Abdulamir, Ahmed Sahib Nordin, Noordiana Yoke, T. S. |
| author_sort | Abu Bakar, Fatimah |
| collection | UPM |
| description | Lactobacillus sp. is probiotic bacteria for which many detection methods were envisaged. However, culture-based methods failed to achieve specific detection of this bacterium due to its presence in mixed bacterial complex communities. The PCR assay was optimized to detect and quantify Lactobacillus sp. specifically in complex microbial community of mixed bacteria. Four DNA extraction methods, DNA integrity, primers specificity and optimized PCR procedure were all tested. It was shown that extracted genomic DNA using Wizard® Genomic DNA Purification Kit showed the highest yield, quality and performance in gel electrophoresis. Moreover, the specificity of the primer set, Lacto-16S-F /Lacto-16S-R, specific for Lactobacillus sp. was checked and found highly specific. In conclusion, the best DNA extraction protocol, working specific primer set and working PCR assay were achieved for achieving efficient, specific and reliable molecular-based, culture-independent, method of detection of lactobacillus sp. in PCR-suppressor highly protein-complex environment of mixed bacteria community. |
| first_indexed | 2024-03-06T07:29:50Z |
| format | Article |
| id | upm.eprints-13937 |
| institution | Universiti Putra Malaysia |
| language | English English |
| last_indexed | 2024-03-06T07:29:50Z |
| publishDate | 2010 |
| publisher | Asian Network for Scientific Information |
| record_format | dspace |
| spelling | upm.eprints-139372015-11-02T06:56:30Z http://psasir.upm.edu.my/id/eprint/13937/ Methods for precise molecular detection of probiotic microflora : using adjusted molecular biology protocols, primer sets and PCR assays. Abu Bakar, Fatimah Abdulamir, Ahmed Sahib Nordin, Noordiana Yoke, T. S. Lactobacillus sp. is probiotic bacteria for which many detection methods were envisaged. However, culture-based methods failed to achieve specific detection of this bacterium due to its presence in mixed bacterial complex communities. The PCR assay was optimized to detect and quantify Lactobacillus sp. specifically in complex microbial community of mixed bacteria. Four DNA extraction methods, DNA integrity, primers specificity and optimized PCR procedure were all tested. It was shown that extracted genomic DNA using Wizard® Genomic DNA Purification Kit showed the highest yield, quality and performance in gel electrophoresis. Moreover, the specificity of the primer set, Lacto-16S-F /Lacto-16S-R, specific for Lactobacillus sp. was checked and found highly specific. In conclusion, the best DNA extraction protocol, working specific primer set and working PCR assay were achieved for achieving efficient, specific and reliable molecular-based, culture-independent, method of detection of lactobacillus sp. in PCR-suppressor highly protein-complex environment of mixed bacteria community. Asian Network for Scientific Information 2010 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/13937/1/Methods%20for%20precise%20molecular%20detection%20of%20probiotic%20microflora.pdf Abu Bakar, Fatimah and Abdulamir, Ahmed Sahib and Nordin, Noordiana and Yoke, T. S. (2010) Methods for precise molecular detection of probiotic microflora : using adjusted molecular biology protocols, primer sets and PCR assays. Biotechnology, 9 (1). pp. 25-32. ISSN 1682-296X 10.3923/biotech.2010.25.32 English |
| spellingShingle | Abu Bakar, Fatimah Abdulamir, Ahmed Sahib Nordin, Noordiana Yoke, T. S. Methods for precise molecular detection of probiotic microflora : using adjusted molecular biology protocols, primer sets and PCR assays. |
| title | Methods for precise molecular detection of probiotic microflora : using adjusted molecular biology protocols, primer sets and PCR assays. |
| title_full | Methods for precise molecular detection of probiotic microflora : using adjusted molecular biology protocols, primer sets and PCR assays. |
| title_fullStr | Methods for precise molecular detection of probiotic microflora : using adjusted molecular biology protocols, primer sets and PCR assays. |
| title_full_unstemmed | Methods for precise molecular detection of probiotic microflora : using adjusted molecular biology protocols, primer sets and PCR assays. |
| title_short | Methods for precise molecular detection of probiotic microflora : using adjusted molecular biology protocols, primer sets and PCR assays. |
| title_sort | methods for precise molecular detection of probiotic microflora using adjusted molecular biology protocols primer sets and pcr assays |
| url | http://psasir.upm.edu.my/id/eprint/13937/1/Methods%20for%20precise%20molecular%20detection%20of%20probiotic%20microflora.pdf |
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