本文選題：脲原體　切入點：血清型　出處：《中國生物制品學雜志》2014年08期 　論文類型：期刊論文

【摘要】：目的制備人脲原體核酸檢測試劑盒國家參考品。方法培養(yǎng)人脲原體兩大生物群14個標準血清型菌株,培養(yǎng)液滅活后經(jīng)顏色改變單位(color change unit,CCU)法與PCR相結合的方法進行定值,組成能覆蓋脲原體所有表型及基因型的14份陽性參考品P1-P14(濃度為106 CCU/ml)及14份檢測限參考品L1-L14(濃度為104 CCU/ml);選擇6份能排除脲原體核酸檢測試劑盒非特異性或交叉反應的樣品組成陰性參考品N1-N6;選擇生物群1的微小脲原體(ureaplasma parvum,UP)1和UP14作為重復性參考品R1、R2(濃度分別為105和104 CCU/ml)。用5家公司生產(chǎn)的脲原體核酸檢測試劑盒對參考品進行驗證,確定準確性、特異性、檢測限、重復性、稀釋線性的性能指標;并考察參考品在不同條件下(2~8℃放置7 d,37℃放置3、7、12 d,-20℃及常溫反復凍融5次)的穩(wěn)定性。結果支原體各培養(yǎng)液經(jīng)CCU法測定,濃度均在106~107 CCU/ml之間,以CCU法測定濃度為靶值,絕對偏差均在±0.5個數(shù)量級以內(nèi)。4家公司生產(chǎn)的不同試劑盒檢測參考品,在準確性、特異性、檢測限及重復性上均符合要求;在稀釋線性上,最低稀釋濃度為102 CCU/ml的樣品只有2家可以檢出,其他幾家的稀釋線性相關系數(shù)以103~106 4個濃度計算,r值均0.990 0;1家(RNA檢測)檢測R2的重復性,CV為5.5%。經(jīng)不同條件處理的參考品的穩(wěn)定性有輕微變化,但均處于設定的可接受范圍內(nèi)。結論制備了人脲原體核酸檢測試劑盒國家參考品,該參考品可滿足人脲原體核酸分型、定性及定量檢測要求,經(jīng)多家實驗室進行驗證,可用于國內(nèi)大多數(shù)試劑盒的性能評價及臨床實驗室質(zhì)量評價。
[Abstract]:Objective to prepare the national reference kit for detection of human Ureaplasma Urealyticum nucleic acid. Methods 14 standard serotypes of two biota of Ureaplasma Urealyticum were cultured. The culture medium was inactivated and determined by the method of color change unit (change) and PCR. 14 positive reference materials covering all phenotypes and genotypes of Ureaplasma Urealyticum P1-P14 (concentration 106 CCU / ml) and 14 limited reference materials L1-L14 (concentration 104 CCU / ml); 6 samples selected to exclude nonspecific or cross reaction of Ureaplasma Urealyticum nucleic acid detection kit. The samples were composed of negative reference material N1-N6; ureaplasma parvum UP1 and UP14 from biome 1 were selected as repeatable reference materials R1UP R2 (concentrations 105 and 104CCU / ml, respectively. The reference was verified with Ureaplasma Urealyticum nucleic acid detection kit produced by five companies, respectively. Determine the accuracy, specificity, detection limit, repeatability, dilution linear performance indicators; The stability of the reference was investigated under different conditions at 8 鈩，

本文編號：1620628