| 摘要: | 結核病是目前普遍存在於世界的疾病，尤其是未開發及開發中國家的慢性傳染病。近年來，結核病似有再次復發流行的趨勢，對於全球的公共衛生是個急迫性的問題。目前結核病的診斷，主要依靠於臨床檢體中培養出Mycobacterium tuberculosis，但是由於Mycobacterium tuberculosis生長緩慢，細菌培養通常需要耗費4~6週的時間，延遲結核病的診斷及治療。近年來，分子鑑定技術的發展應用，即可大符縮短檢測時間，廣泛使用於分枝桿菌屬的診斷。
本研究自2011年11月至2013年4月，由37病患總共收集108個痰液檢體，將所收集到的病患檢體，經消化去污染後，一方面進行分枝桿菌屬的培養及抗酸性染色，同時將檢體進行 mRNA以及DNA的抽取，將mRNA先反轉錄成cDNA，使用Cobas TaqMan MTB test，利用Real time PCR的方法，偵測結核分枝桿菌群（Mycobacterium tuberculosis complex），將所得結果分別與培養結果作比較。研究結果DNA Real time PCR 此方法的Sensitivity、Specificity、Positive predictive value（PPV）、Negative predictive value（NPV）、Prevalence及Efficiency分別為90.00%、79.17%、84.38%、86.36%、55.56%及85.19%。mRNA RT-Real time PCR此方法的Sensitivity、Specificity、Positive predictive value（PPV）、Negative predictive value（NPV）、Prevalence及Efficiency分別為96.67%、79.17%、85.29%、95.00%、55.56%和88.89%。根據統計結果顯示mRNA RT-Real time PCR比DNA Real time PCR有較高的敏感性，與細菌培養的結果一致性也較高(88.89% V.S. 85.19%)。所以此次研究的結論，mRNA RT-Real time PCR能更有效縮短鑑定時間且準確地從臨床檢體檢測Mycobacterium tuberculosis complex，能夠作為一個有效的分子鑑定工具。
Tuberculosis remains a highly prevalent disease in the world, especially in the undeveloped and developing countries. The resurgence of tuberculosis has created a global public health emergency. Currently, the diagnosis of tuberculosis mainly relies on cultures of Mycobacterium tuberculosis from clinical specimens. However, the diagnosis and treatment of tuberculosis are often delayed due to the slow growth of Mycobacterium tuberculosis, which usually takes 4 to 6 weeks. In recent years, the development and application of molecular identification techniques allow operators to timely detect Mycobacterium.
This study collected 108 sputum samples from 37 patients. After digesting, decontaminating and neutralization the specimen, we performed the Mycobacterial culture and acid-fast staining. On the other hand, the mRNA and DNA were extracted from decontaminated specimens. The mRNA then was reverse transcribed into cDNA. We used a real-time PCR assay for M. tuberculosis complex, the Cobas TaqMan MTB test (Roche Diagnostics, Basel,Switzerland). Finally, the results were compared to culture findings. The overall Sensitivity, Specificity, Positive predictive values (PPV), Negative predictive values (NPV), Prevalence and Efficiency were 90.00%, 79.17%, 84.38% 86.36%, 55.56% and 85.19%, respectively, for DNA Real time PCR, and 96.67%, 79.17%, 85.29% 95.00%, 55.56%and 88.89%, respectively, for mRNA RT-Real time PCR. Furthermore, the mRNA RT-Real time PCR showed a higher sensitivity than DNA Real time PCR. The overall agreement between the culture and mRNA RT-Real time PCR results was 88.89%, which was significantly better than between the culture and DNA Real time PCR results (88.89% V.S. 85.19%). We concluded that the mRNA RT-Real time PCR is a useful molecular identification technique to shorten the detection time and provide accurate identification of M. tuberculosis complex directly from clinical specimens. |
