实用医学杂志2012，Vol.28Issue(12)：2067-2069,3.DOI:10.3969/j.issn.1006-5725.2012.12.054
荧光定量PCR检测抗酸染色涂片刮削物中结核分枝杆菌DNA的可行性
The feasibility of detecting mycobacterium tuberculosis DNA from scraping things of acid-fast stained smear slides by fluorescence quantitative PCR
摘要
Abstract
Objective To analyze the feasibility of detecting Mycobacterium tuberculosis DNA from scraping things of acid-fast stained smear slides by fluorescence quantitative PCR. Methods 281 acid-fast-stained smear slides [including 80 negative slides marked as -, 25 slides with a score of 1-9 bacteria per 300 microscope fields (marked as +), 95 slides with a score of 1 + , 40 slides with a score of 2+, 21 slides with a score of 3+, and 20 slides with a score of 4+ according to acid-fast-stained smear microscopy procedures] were collected, and their mycobacterium tuberculosis DNA from scraping things of stained smear slides were detected by fluorescence quantitative PCR. Then the detecting results were analyzed in accordance with positive degree of acid-fast-stained smear, and the correlation between positive degree of acid-fast stained smear and target DNA copies was studied by Spearman rank correlation. Results Of these scraping things of stained smear slides from ± slides to 4+ slides in order, 7, 48, 31, 19, and 20 cases were positive respectively for mycobacterium tuberculosis DNA detection. The positive rates were 28.00%, 50.53%, 77.50%, 90.48%, and 100.00%, respectively, whilst it was 62.19% for all these samples. A positive correlation between positive degree of acid-fast-stained smear and target DNA copies was found (r = 0.684, P ＜ 0.001). As for scraping things of 80 negative slides, none of them was positive for mycobacterium tuberculosis DNA detection. Conclusion Fluorescence quantitative PCR is effective in detecting mycobacterium tuberculosis DNA from scraping things of acid-fast stained smear slides above 2+. But for smear with a score of 1 +, it should be studied further why presented very lower positive rate than theory prediction as bacteria number in it is within the sensitivity range of fluorescence quantitative PCR technique theoretically.关键词
荧光定量PCR/涂片/结核分枝杆菌/抗酸染色Key words
Fluorescence quantitative PCR/ Smear/ Mycobacterium tuberculosis/ Acid-fast staining引用本文复制引用
谭守勇,刘志辉,邓向斌..荧光定量PCR检测抗酸染色涂片刮削物中结核分枝杆菌DNA的可行性[J].实用医学杂志,2012,28(12):2067-2069,3.基金项目
国家"十一.五"重大科技专项资助项目(编号:2009ZX10003-018) （编号:2009ZX10003-018）
广州市医药卫生科技项目重点专项(编号:2009-Zdi-17) （编号:2009-Zdi-17）
