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Cell Journal، جلد ۲۱، شماره ۱، صفحات ۴۳-۴۸
عنوان فارسی
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عنوان انگلیسی Detection of Mycoplasma Contamination of Cell Culture by A Loop-Mediated Isothermal Amplification Method
چکیده انگلیسی مقاله Objective: Mycoplasmas being the most serious contaminants of cell culture and affect biological and diagnostic tests in cell culture. Mycoplasma diagnosis is conducted on the basis of culture and molecular methods. These methods are different from each other in terms of accuracy, reliably and sensitivity. Loop-mediated isothermal amplification (LAMP) amplify target DNA in a highly specific and rapid manner. The aim of this study was designing a LAMP method for the rapid detection of Mycoplasma in culture samples. Materials and methods: In this descriptive laboratory study for the LAMP detection of Mycoplasma contaminations in cell culture, we used specifically designed primers to target the 16S rRNA conserved gene of Mycoplasma spp. For a positive control structure, 16S rRNA amplified based on PCR was cloned in plasmid vector and then sequenced. The assay specificity was evaluated using Mycoplasma genomic DNA and a panel containing genomes of gram-positive and gram-negative organisms. Results: Our study defines a rapid, sensitive and cost-effective LAMP method for Mycoplasma contamination of cell cultures. The results demonstrate that it has high specificity (100%) for the amplification of Mycoplasma strains. This method provides advantages such as high speed (multiplication within 60 min), no need to expensive laboratory equipment compared to PCR-based detection. Conclusion: Our study is the first report about application of LAMP assay based on 16S rRNA gene for determination of Mycoplasma strains and considered as useful tool for the rapid diagnosis in laboratory cell culture
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نویسندگان مقاله | Zohre Soheily


| Mohammad Soleimani


| Keivan Majidzadeh-Ardebili


نشانی اینترنتی http://celljournal.org/journal/article/abstract/5624
فایل مقاله اشکال در دسترسی به فایل - ./files/site1/rds_journals/16/article-16-1043612.pdf
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