Evaluation of Different Nucleic Acid Extraction Methods from Neisseria Gonorrhoeae_West China Medical Journal

Authors：

YING Bin-wu. Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P. R. China;

Corresponding author：

Keywords：

淋病奈瑟菌; DNA提取; 实时荧光定量聚合酶链反应

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目的　比较评价磁珠法、柱子吸附法和煮沸裂解法3种不同的核酸提取方法对淋病奈瑟菌（淋球菌）DNA提取效率的差异。方法　对已确诊的淋球菌培养基挑取20个小菌落制成悬液，做1︰1～1︰100 000稀释度稀释，分别使用磁珠法、柱子吸附法和煮沸裂解法进行DNA提取，用实时荧光定量PCR对其提取效率进行评价。　结果　在不同浓度淋球菌的DNA提取中，以磁珠法的敏感性和重复性最高，而柱子吸附法和煮沸裂解法次之，＜1×105 copies /mL以下的拷贝数，3种DNA提取方法之间重复性和敏感性差异明显，磁珠法的提取效率最高，柱子吸附法次之，煮沸裂解法最差；＞1×105 copies/mL的细菌拷贝数的样本，3种方法差异随浓度增高不断缩小。结论　磁珠法提取淋球菌DNA的效率、敏感性和重复性最高，在实际临床科研工作中，各实验室可根据自身实际情况选择不同的DNA提取方法。

Citation： ZHUANG Jie,LIU Xianzhong,YE Yuanxin,ZHOU Yi,SONG Xingbo,LU Xiaojun,WANG Jun,DING Liu,DAI Bo. Evaluation of Different Nucleic Acid Extraction Methods from Neisseria Gonorrhoeae. West China Medical Journal, 2012, 27(4): 560-563. doi: Copy

1.	Mackay IM. Real-time PCR in themicrobiology laboratory[J]. Clin Microbiol Infect, 2004, 10(3): 190-212.
2.	姚远, 张国富, 刘玉英. PCR污染与对策及假阴性原因[J]. 实用医技杂志, 2002, 9(4): 276-277.
3.	Knepp JH., Geahr MA, Forman MS. Comparison of automated and manual nucleic acid extraction methods for detection of enterovirus RNA[J]. J Clin Microbiol, 2003, 41(8): 3532-3536.
4.	Konomi N, Lebwohl E, Zhang D. Comparison of DNA and RNA extraction methods for mummified tissues[J]. Mol Cell Probes, 2002, 16(6): 445-451.
5.	Sweet D, Lorente M, Valenzuela A, et al. Increasing DNA extraction yield from saliva stains with a modified Chelex method[J]. Forensic Sci Int, 1996, 83(3): 167-177.
6.	Stein A, Raoult D. A simple method amplification of DNA from paraffin embedded tissues[J]. Nucleic Acids Res, 1992, 20(19): 5237-5238.
7.	Dundas N, Leos NK, Mitui M, et al. Comparison of automated nucleic acid extraction methods with manual extraction[J]. J Mol Diagn, 2008, 10(4): 311-316.
8.	Claassen M, Van zyl GU, Preiser W. Extraction buffer contaminated bacterially as a cause of invalid HIV-1 viral load results on the NucliSens EasyQ system[J]. J Virol Methods, 2008, 150(12): 80-81.
9.	Stevens W, Horsfield P, Scott LE. Evaluation of the performance of the automated NucliSENS easyMAG and EasyQ systems versus the Roche AmpliPrep-AMPLICOR combination for high-throughput monitoring of human immunodeficiency virus load[J]. J Clin Microbiol, 2007, 45(4): 1244-1249.
10.	Thakur R, Sarma S, Goyal R. Comparison of DNA extraction protocols for mycobacterium tuberculosis in diagnosis of tuberculous meningitis by real-time polymerase chain reaction[J]. J Glob Infect Dis, 2011, 3(4): 353-356.
11.	Ursi D, Dirven K, Loens K, et al. Detection of mycoplasma pneumoniae in respiratory samples by real-time PCR using an inhibition control[J]. Microbiol Methods, 2003, 55(1): 149-153.
12.	Bahl MI, Bergström A, Licht TR. Freezing fecal samples prior to DNA extraction affects the Firmicutes to Bacteroidetes ratio determined by downstream quantitative PCR analysis[J]. FEMS Microbiol Lett, 2012, 329(2): 193-197.
13.	Kouduka M, Suko T, Morono Y, et al. A new DNA extraction method by controlled alkaline treatments from consolidated subsurface sediments[J]. FEMS Microbiol Lett, 2012, 326(1): 47-54.

1. 　Mackay IM. Real-time PCR in themicrobiology laboratory[J]. Clin Microbiol Infect, 2004, 10(3): 190-212.
2. 　姚远, 张国富, 刘玉英. PCR污染与对策及假阴性原因[J]. 实用医技杂志, 2002, 9(4): 276-277.
3. 　Knepp JH., Geahr MA, Forman MS. Comparison of automated and manual nucleic acid extraction methods for detection of enterovirus RNA[J]. J Clin Microbiol, 2003, 41(8): 3532-3536.
4. 　Konomi N, Lebwohl E, Zhang D. Comparison of DNA and RNA extraction methods for mummified tissues[J]. Mol Cell Probes, 2002, 16(6): 445-451.
5. 　Sweet D, Lorente M, Valenzuela A, et al. Increasing DNA extraction yield from saliva stains with a modified Chelex method[J]. Forensic Sci Int, 1996, 83(3): 167-177.
6. 　Stein A, Raoult D. A simple method amplification of DNA from paraffin embedded tissues[J]. Nucleic Acids Res, 1992, 20(19): 5237-5238.
7. 　Dundas N, Leos NK, Mitui M, et al. Comparison of automated nucleic acid extraction methods with manual extraction[J]. J Mol Diagn, 2008, 10(4): 311-316.
8. 　Claassen M, Van zyl GU, Preiser W. Extraction buffer contaminated bacterially as a cause of invalid HIV-1 viral load results on the NucliSens EasyQ system[J]. J Virol Methods, 2008, 150(12): 80-81.
9. 　Stevens W, Horsfield P, Scott LE. Evaluation of the performance of the automated NucliSENS easyMAG and EasyQ systems versus the Roche AmpliPrep-AMPLICOR combination for high-throughput monitoring of human immunodeficiency virus load[J]. J Clin Microbiol, 2007, 45(4): 1244-1249.
10. 　Thakur R, Sarma S, Goyal R. Comparison of DNA extraction protocols for mycobacterium tuberculosis in diagnosis of tuberculous meningitis by real-time polymerase chain reaction[J]. J Glob Infect Dis, 2011, 3(4): 353-356.
11. 　Ursi D, Dirven K, Loens K, et al. Detection of mycoplasma pneumoniae in respiratory samples by real-time PCR using an inhibition control[J]. Microbiol Methods, 2003, 55(1): 149-153.
12. 　Bahl MI, Bergström A, Licht TR. Freezing fecal samples prior to DNA extraction affects the Firmicutes to Bacteroidetes ratio determined by downstream quantitative PCR analysis[J]. FEMS Microbiol Lett, 2012, 329(2): 193-197.
13. 　Kouduka M, Suko T, Morono Y, et al. A new DNA extraction method by controlled alkaline treatments from consolidated subsurface sediments[J]. FEMS Microbiol Lett, 2012, 326(1): 47-54.

West China Medical Journal

Evaluation of Different Nucleic Acid Extraction Methods from Neisseria Gonorrhoeae

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