| 中文摘要 |
本實驗乃是以臨床檢體及CAP能力試驗檢體來評估TIB MOLBIOL LightMix(R) Kit HSV-1/2試劑於LightCycler(R) 480 II real-time PCR系統檢測之效能，並與原使用之semi-nested PCR方法做一比較。以108支臨床病人之剩餘檢體進行HSV real-time qPCR與傳統semi-nested PCR一致性之評估，二種方法總相同率為96.3 %，Kappa檢定之k值為0.911。比較real-time qPCR及semi-nested PCR的靈敏度，結果分別可測得10 TCID50及0.1 TCID50。由於LightMix(R) Kit HSV-1/2可提供定量檢測結果，故另評估其最低偵測極限、再現性及線性。以試劑組內附的標準品測試最低偵測極限之結果為10 copies / reaction，與試劑說明書相符。再現性的評估為計算標準品（102 to 106 copies / reaction）log值的累計CV值，結果皆小於5.0 %。迴歸線性分析R2值為0.983，線性良好。
The purpose of this study was to evaluate the performance of TIB MOLBIOL LightMix(R) KitHSV-1/2 on LightCycler(R) 480 II real-time qPCR system, and compare the results with a routinelyused in-house semi-nested PCR. The clinical and proficiency testing samples of College of AmericanPathology (CAP) were used. A total of 108 clinical samples were in parallel tested by real-timeqPCR and semi-nested PCR. Results indicated the total agreement between the two methods was96.3%, with Kappa coefficient (k) being 0.911. The sensitivity of real-time qPCR and semi-nestedPCR were also tested in triplicate using tenfold serially diluted HSV-1, and the results were 10TCID50 and 0.1 TCID50, respectively. Since the LightMix(R) Kit HSV-1/2 kit can provide quantitativeresults, we also evaluated the lowest limit of detection (LOD), repeatability and linearity of the kit.The LOD evaluated with standards provided in the kit was 10 copies / reaction, as indicated in thepackage insert. The repeatability of the kit was evaluated with cumulated Coefficient of variation(CV) of the log transformed data of standards (from 102 to 106 copies / reaction). Results shownwere all below 5.0 %. Furthermore, R-squared value of the linear regression analysis was 0.983,showing excellent linearity. Briefly, our study indicated the TIB MOLBIOL LightMix(R) KitHSV-1/2 combined with LightCycler(R) 480 II real-time qPCR system is a good method for detectionof HSV. |
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