Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction
Background: Nontuberculous mycobacteria (NTM) are a group of opportunistic pathogens and these are widely dispersed in water and soil resources. Identification of mycobacteria isolates by conventional methods including biochemical tests, growth rates, colony pigmentation, and presence of acid-fast b...
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| Format: | Article |
| Language: | English |
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Wolters Kluwer Medknow Publications
2017-01-01
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| Series: | Advanced Biomedical Research |
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| Online Access: | http://www.advbiores.net/article.asp?issn=2277-9175;year=2017;volume=6;issue=1;spage=133;epage=133;aulast=Mohammadi |
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| author | Samira Mohammadi Bahram Nasr Esfahani Sharareh Moghim Hossein Mirhendi Fatemeh Riyahi Zaniani Hajieh Ghasemian Safaei Hossein Fazeli Mahshid Salehi |
| author_facet | Samira Mohammadi Bahram Nasr Esfahani Sharareh Moghim Hossein Mirhendi Fatemeh Riyahi Zaniani Hajieh Ghasemian Safaei Hossein Fazeli Mahshid Salehi |
| author_sort | Samira Mohammadi |
| collection | DOAJ |
| description | Background: Nontuberculous mycobacteria (NTM) are a group of opportunistic pathogens and these are widely dispersed in water and soil resources. Identification of mycobacteria isolates by conventional methods including biochemical tests, growth rates, colony pigmentation, and presence of acid-fast bacilli is widely used, but these methods are time-consuming, labor-intensive, and may sometimes remain inconclusive. Materials and Methods: The DNA was extracted from NTM cultures using CTAB, Chelex, Chelex + Nonidet P-40, FTA® Elute card, and boiling The quantity and quality of the DNA extracted via these methods were determined using UV-photometer at 260 and 280 nm, and polymerase chain reaction (PCR) amplification of the heat-shock protein 65 gene with serially diluted DNA samples. Results: The CTAB method showed more positive results at 1:10–1:100,000 at which the DNA amount was substantial. With the Chelex method of DNA extraction, PCR amplification was detected at 1:10 and 1:1000 dilutions. Conclusions: According to the electrophoresis results, the CTAB and Chelex DNA extraction methods were more successful in comparison with the others as regard producing suitable concentrations of DNA with the minimum use of PCR inhibitor. |
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| id | doaj.art-4fe2b708abcb4720a9be157d5b0b0208 |
| institution | Directory Open Access Journal |
| issn | 2277-9175 |
| language | English |
| last_indexed | 2024-12-11T11:20:21Z |
| publishDate | 2017-01-01 |
| publisher | Wolters Kluwer Medknow Publications |
| record_format | Article |
| series | Advanced Biomedical Research |
| spelling | doaj.art-4fe2b708abcb4720a9be157d5b0b02082022-12-22T01:09:12ZengWolters Kluwer Medknow PublicationsAdvanced Biomedical Research2277-91752017-01-016113313310.4103/2277-9175.217216Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain ReactionSamira MohammadiBahram Nasr EsfahaniSharareh MoghimHossein MirhendiFatemeh Riyahi ZanianiHajieh Ghasemian SafaeiHossein FazeliMahshid SalehiBackground: Nontuberculous mycobacteria (NTM) are a group of opportunistic pathogens and these are widely dispersed in water and soil resources. Identification of mycobacteria isolates by conventional methods including biochemical tests, growth rates, colony pigmentation, and presence of acid-fast bacilli is widely used, but these methods are time-consuming, labor-intensive, and may sometimes remain inconclusive. Materials and Methods: The DNA was extracted from NTM cultures using CTAB, Chelex, Chelex + Nonidet P-40, FTA® Elute card, and boiling The quantity and quality of the DNA extracted via these methods were determined using UV-photometer at 260 and 280 nm, and polymerase chain reaction (PCR) amplification of the heat-shock protein 65 gene with serially diluted DNA samples. Results: The CTAB method showed more positive results at 1:10–1:100,000 at which the DNA amount was substantial. With the Chelex method of DNA extraction, PCR amplification was detected at 1:10 and 1:1000 dilutions. Conclusions: According to the electrophoresis results, the CTAB and Chelex DNA extraction methods were more successful in comparison with the others as regard producing suitable concentrations of DNA with the minimum use of PCR inhibitor.http://www.advbiores.net/article.asp?issn=2277-9175;year=2017;volume=6;issue=1;spage=133;epage=133;aulast=MohammadiDNA extraction methodsheat-shock protein 65nontuberculous mycobacteriapolymerase chain reaction |
| spellingShingle | Samira Mohammadi Bahram Nasr Esfahani Sharareh Moghim Hossein Mirhendi Fatemeh Riyahi Zaniani Hajieh Ghasemian Safaei Hossein Fazeli Mahshid Salehi Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction Advanced Biomedical Research DNA extraction methods heat-shock protein 65 nontuberculous mycobacteria polymerase chain reaction |
| title | Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction |
| title_full | Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction |
| title_fullStr | Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction |
| title_full_unstemmed | Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction |
| title_short | Optimal DNA Isolation Method for Detection of Nontuberculous Mycobacteria by Polymerase Chain Reaction |
| title_sort | optimal dna isolation method for detection of nontuberculous mycobacteria by polymerase chain reaction |
| topic | DNA extraction methods heat-shock protein 65 nontuberculous mycobacteria polymerase chain reaction |
| url | http://www.advbiores.net/article.asp?issn=2277-9175;year=2017;volume=6;issue=1;spage=133;epage=133;aulast=Mohammadi |
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