A rapid and simple method for identifying Mycobacterium tuberculosis W-Beijing strains based on detection of a unique mutation in Rv0927c by PCR-SSCP |
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| Institution: | 1. Shanghai University of Traditional Chinese Medicine, Shanghai 201203, People''s Republic of China;2. State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Science, Fudan University, Shanghai 200433, People''s Republic of China;3. Shanghai First People''s Hospital, Shanghai Jiao Tong University, Shanghai 200080, People''s Republic of China;4. Huashan Hospital, Fudan University, Shanghai 200433, People''s Republic of China;5. Molecular Pathogenesis Centre, University of Lisbon, School of Pharmacy, Lisbon, Portugal;6. Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, Shanghai 200032, People''s Republic of China;7. Department of Molecular Microbiology & Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD 21205, USA;1. Research Department of Plant Biology and Biotechnology, Loyola College, Nungambakkam, Chennai, Tamil Nadu, India;2. Facultad de Medicina Veterinaria y Zootecnia, Universidad Autónoma del Estado de México, Toluca, Mexico;3. Facultad de Medicina Veterinaria y Zootecnia, Universidad Autónoma de Guerrero, Cd. Altamirano, Guerrero, México;1. Institute for Anatomy and Cell Biology, German Center for Lung Research, Justus Liebig University, 35385 Giessen, Germany;2. Department of Pharmacology, Max Planck Institute for Heart and Lung Research, 61231 Bad Nauheim, Germany;3. Department of Medicine III, University Medical Center Hamburg-Eppendorf, 20251 Hamburg, Germany;4. Center for Integrative Physiology and Molecular Medicine, Saarland University, 66421 Homburg, Germany;5. Institute of Anatomy and Cell Biology, Philipps-University, 35037 Marburg, Germany;6. Institute of Biochemistry, Justus Liebig University, 35385 Giessen, Germany;7. Experimental Pharmacology, Center for Molecular Signaling (PZMS), Saarland University, 66421 Homburg, Germany;8. Institute of Medical Microbiology, German Center for Infection Research, Justus Liebig University, 35385 Giessen, Germany;9. Institute of Hygiene and Infectious Diseases of Animals, Justus Liebig University, 35392 Giessen, Germany;10. Center for Interstitial and Rare Lung Diseases, Department of Internal Medicine, German Center for Lung Research, Justus Liebig University, 35385 Giessen, Germany;11. Agaplesion Lung Clinic Waldhof-Elgershausen, 35753 Greifenstein, Germany;12. Walther-Straub Institute of Pharmacology and Toxicology, German Center for Lung Research, Ludwig-Maximilians-Universität München, 80336 Munich, Germany;13. Institute of Physiology and Pathophysiology, Philipps-University, 35037 Marburg, Germany;14. Department of Pharmacology and Clinical Pharmacy, FB14, Goethe University Frankfurt, 60438 Frankfurt am Main, Germany;15. Laboratory of Cell Calcium Signaling, Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, 201210 Shanghai, China;16. Department of Informatics and Microsystems Technology, University of Applied Sciences, 67659 Kaiserslautern, Germany;1. The McDonnell Genome Institute, Washington University, St. Louis, MO 63108, USA;2. Department of Genetics, Washington University, St. Louis, MO 63108, USA;3. Siteman Cancer Center, Washington University, St. Louis, MO 63108, USA;4. Department of Medicine, Washington University, St. Louis, MO 63108, USA;5. Department of Mathematics, Washington University, St. Louis, MO 63108, USA;6. Department of Pathology and Immunology, Washington University, St. Louis, MO 63108, USA;7. Department of Pediatrics, Division of Hematology/Oncology, Washington University, St. Louis, MO 63108, USA;8. Department of Biomedical Engineering, Washington University, St. Louis, MO 63108, USA;1. Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX, United States;2. Department of Pediatrics, University of Texas Medical Branch, Galveston, TX, United States;3. WHO Collaborating Center, University of Texas Medical Branch, Galveston, TX, United States;1. State Key Laboratory of Molecular Development Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China;2. University of Chinese Academy of Sciences, Beijing 100101, China;3. Department of Pathology, Stanford University School of Medicine, Stanford, CA 94305, USA;4. Department of Endocrinology and Metabolism, Huashan Hospital, Fudan University, Shanghai 200040, China;5. Department of Obstetrics and Gynecology, Baoding Second Central Hospital, Baoding, Hebei 072750, China;6. Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Beijing 100101, China;7. Chinese Institute for Brain Research, Beijing 102206, China;8. Beijing Children''s Hospital, Capital Medical University, Beijing 100045, China |
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| Abstract: | Recently we have found that W-Beijing Mycobacterium tuberculosis strains have a unique in-frame trinucleotide (AGC) deletion at position 421 of Rv0927c and a −127G → A mutation in Rv0927c-pstS3 intergenic region. Based on detecting the 421 trinucleotide deletion of these two mutations which can alter the ssDNA conformation more extensively than the other, we developed a PCR-SSCP method for rapid identification of W-Beijing strains among non-Beijing strains. Altogether, 104 clinical isolates were analyzed, including 68 W-Beijing strains and 36 non-Beijing strains. We found that PCR-SSCP successfully differentiated all the W-Beijing strains from the non-Beijing strains. In addition, we unexpectedly discovered that SDS-PAGE protein gels had better resolving power than conventional TBE polyacrylamide gel in detecting the AGC deletion mutation in the SSCP analysis. |
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