科研動(dòng)態(tài)

為填補(bǔ)測(cè)量不確定度在獸醫(yī)領(lǐng)域病原學(xué)檢測(cè)的空白，建立豬繁殖與呼吸綜合征病毒（PRRSV）實(shí)時(shí)熒光定量RT-PCR檢測(cè)試劑盒測(cè)量不確定度評(píng)估模型，對(duì)市售的3種美洲型PRRSV實(shí)時(shí)熒光RT-PCR檢測(cè)試劑盒進(jìn)行了測(cè)量不確定度評(píng)估，評(píng)估了移液器、溫度、重復(fù)性測(cè)量因素對(duì)試驗(yàn)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(cè)量對(duì)試驗(yàn)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(cè)量不確定度模型縮小了試劑盒對(duì)可疑樣品的判定區(qū)域，對(duì)樣品臨界參考值的檢測(cè)結(jié)果判定具有實(shí)際意義。通過(guò)比對(duì)4個(gè)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(shí)時(shí)熒光RT-PCR定量檢測(cè)試劑盒測(cè)量不確定度模型為獸醫(yī)實(shí)驗(yàn)室檢測(cè)結(jié)果不確定度評(píng)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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