為填補(bǔ)測量不確定度在獸醫(yī)領(lǐng)域病原學(xué)檢測的空白，建立豬繁殖與呼吸綜合征病毒（PRRSV）實時熒光定量RT-PCR檢測試劑盒測量不確定度評估模型，對市售的3種美洲型PRRSV實時熒光RT-PCR檢測試劑盒進(jìn)行了測量不確定度評估，評估了移液器、溫度、重復(fù)性測量因素對試驗結(jié)果產(chǎn)生的不確定度影響。研究數(shù)據(jù)表明：移液器與重復(fù)性測量對試驗結(jié)果產(chǎn)生的不確定度影響較大，應(yīng)予以重視，需規(guī)范操作；建立的3種試劑盒測量不確定度模型縮小了試劑盒對可疑樣品的判定區(qū)域，對樣品臨界參考值的檢測結(jié)果判定具有實際意義。通過比對4個廠家的核酸提取試劑，篩選出了提取效率最高的試劑盒。本研究建立的PRRSV實時熒光RT-PCR定量檢測試劑盒測量不確定度模型為獸醫(yī)實驗室檢測結(jié)果不確定度評估及應(yīng)用奠定了基礎(chǔ)。
Study on the Uncertainty of Real-time PCR Kits for Quantitative Detection of PRRSV of American Type
In order to fill in the blank of the measurement uncertainty in the field of veterinary etiology，an evaluation model of measurement uncertainty for real-time PCR kits for quantitative detection of PRRSV was established，and three kinds of commercial RT-PCR kits（detecting PRRSV strain of American type）were evaluated，including the influence of pipette，temperature and repeated measurement on test results. It was found that the test results were greatly influenced by the pipette and repeated measurement，which should be paid attention to，and the operations should be standardized；the determination area of the kits for suspected samples was reduced，which contributed to the determination of the critical reference value of samples. The kit with highest extraction efficiency was selected through comparing the nucleic acid extraction reagents produced by four manufacturers. In conclusion，a foundation was hereby laid for evaluation and application of measurement uncertainty in veterinary laboratories by the model established in this study.
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