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《Acta Academiae Medicine Militaris Tertiae》 2002-01
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Influential factors of strand displacement amplification system with commercial reagents

JIANG Tian lun, FU Wei ling,ZHANG Bo,CHEN Ming,ZHANG Xue,ZHAO Shu ming (Department of Clinical Laboratories, Southwest Hospital, Third Military Medical University, Chongqing 400038, China)  
Objective To explore the effects of the restriction endonuclease, polymerase, buffer system, BSA and DTT on strand displacement amplification (SDA) reaction and establish SDA reaction system with commercial reagents. Methods The Mycobacterium tuberculosis IS6110 segment was used as the detecting target and the corresponding primers were designed, experiment SDA system was finally established with commercial BsoB Ⅰ and exo Bst (both from New England Biolabs) as SDA enzymes. The influences of the restriction endonuclease and the polymerase on SDA were obtained by comparing the brightness of the electrophoresis bands of SDA products in the systems with different BsoB Ⅰ and exo Bst concentrations. The effects of the buffers on SDA were explored through the comparison of the SDA products with electrophoresis in phosphate buffer (pH=7.6) and the Tris HCl buffers (including NE Buffer2, ThermoPol Buffer and EcoPol Buffer). Electrophoresis was also used to study SDA with DTT or without. Results The concentration of BsoB Ⅰ had a strong effect to the SDA rate, but in a certain concentration range. While, exo Bst concentration had no effect in this event. SDA reactions worked well in ThermoPol Buffer. The DTT was observed having no effect on SDA reaction. Conclusion The results show that SDA works well with commercial available reagents.
【Fund】: 国家“九五”科技攻关项目 ( 96 A2 3 0 4 0 4 );; 国家自然科学基金资助面上项目 ( 39870 831 );; 国家高技术计划生物技术领域青年基金资助项目 (“86 3”项目 ;; 全军“九五”医学科研计划资助面上项目( 98MO94 ;; 全军“十五”医学科研计划资助面上项目 ( 0 1MA1
【CateGory Index】: R346
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【References】
Chinese Journal Full-text Database 1 Hits
1 CHEN Bin, FU Wei-ling, JIANG Tian-lun,HUANG Jun-fu (Department of Laboratory Medicine, Southwest Hospital, the Third Military Medical University, Chongqing 400038, China);Detection of Pseudomonas aeruginosaby SDA Piezoelectric DNA Sensors Array: A Clinical Application[J];Chinese Journal of Nosoconmiology;2004-01
【Co-references】
Chinese Journal Full-text Database 8 Hits
1 JIANG Tian lun, FU Wei ling,ZHANG Bo, CHEN Ming, LIU Ming hua, ZHANG Xue, ZHAO Shu ming, YU Fan, CHEN Xiao bin (Department of Clinical Laboratories, Southwest Hospital, Third Military Medical University, Chongqing 400038, China);Strand displacement amplification piezoelectric DNA sensor[J];Acta Academiae Medicine Militaris Tertiae;2002-01
2 GUO Yan Hai 1, ZHONG Yong Mei 2, ZHAO Jin Rong 1, CUI Da Xiang 1, YAN Xiao Jun 1 1Institute of Genic Diagnosis of Chinese PLA, Fourth Military Medical University, Xi'an 710033, China, 2Department of Laboratory, Jinjiang Hospital, Fujian, Jinjian 362200, China;Development and evaluation of a PCR-ELISA method for detecting TB DNA and application of it in testing against samples[J];Journal of The Fourth Military Medical University;2002-22
3 LIAO Jin liang, TAN Yao ju, CHEN Zhi cheng, et al. Guangzhou Chest Hospital, Guangzhou 510095;Detection of DNA of Mycobacterium tuberculosis in cerebrospinal fluid by Amplisensor-PCR[J];Buccetin of Chinese Antituberculosis Assoclation;2001-04
4 National Technical Steering Group of the Epidemiological Sampling Survey for Tuberculosis Office of the Nationwide Epidemiological Sampling Survey for Tuberculosis;Report on nationwide random survey for the epidemiology of tuberculosis in 2000[J];Buccetin of Chinese Antituberculosis Assoclation;2002-02
5 LI Guo li,ZHAO Ming,Wang Wei,et al.The 309th Hospital of PLA, Beijing 100091;Clinical value of fluorogenic probe hybridzation assay for detecting Mycobacterium tuberculosis in clinical specimens[J];Bulletin of Chinese Antituberculosis Association;2003-02
6 HE Min *, ZENG Er-liang, ZHENG Yan-yan, TANG Zhuo, LU Xiang-chan, SUN Bi-hui, XU Ding-kong, ZHANG Zhi-yong, YANG Li. *Medical Scientific Research Center, Guangxi Medical University, Nanning 530021, China;Identification of Mycobacterium tuberculosis and rifampin-resistant strains by gene-chips[J];Chinese Journal of Epidemiology;2003-05
7 LI Guoli, ZHAO Ming, ZHUANG Yuhui, et al. 309th Hospital, Beijing 100091, China;PCR-molecular beacon assay for detecting Mycobacterium tuberculosis in clinical specimens[J];Chinese Journal of Laboratory Medicine;2001-02
8 CHEN Ming,FU Wei ling,LIU Ming hua,ZHANG Bo,JIANG Tian lun,GONG Yan wen,CAI Guo ru (Department of Laboratory Diagnosis, Southwest Hospital, the Third Military Medical University, Chongqing 400038, China);Sensitivity Improvement of Detecting MRSA Using Gene Sensor by Strand Extension Reaction with Long and Short Chain Probes[J];Chinese Journal of Nosoconmiology;2003-03
【Secondary References】
Chinese Journal Full-text Database 3 Hits
1 SUN Hai-bai,ZHANG Li-xia,ZHANG Li;Comparisons of diagnostic and curative effects between the TB anti-body titer and the specificity rates of sputum culture[J];Journal of Clinical Pulmonary Medicine;2008-08
2 CHEN Bin, FU Wei-ling, HUANG Qing, ZHANG Xiao-bing, HUANG Jun-fu(Southwest Hospital,the Third Military Medical University,Chongqing 400038, China);Detection of Pseudomonas aeruginosa by Piezoelectric Immunosensor Microarray: An Experimental Research[J];Chinese Journal of Nosoconmiology;2005-04
3 WANG Feng~*,FU Wei-ling,XU Xue-qing,XIA Han,LUO Yang(*Southwest Hospital,the Third Military Medical University,Chongqing,400038,China);Piezoelectric Gene Biosensors for Detection of Pseudomonas aeruginosa Using Signal Amplification Method with Cationic Conjugated Polymers[J];Chinese Journal of Nosocomiology;2006-11
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