国际输血及血液学杂志
國際輸血及血液學雜誌
국제수혈급혈액학잡지
INTERNATIONAL JOURNAL OF BLOOD TRANSFUSION AND HEMATOLOGY
2012年
5期
385-389
,共5页
金士正%甄建新%何柳媚%徐筠娉%邓志辉
金士正%甄建新%何柳媚%徐筠娉%鄧誌輝
금사정%견건신%하류미%서균빙%산지휘
杀伤细胞免疫球蛋白样受体%流式序列特异性寡核苷酸探针%序列特异性引物-PCR%KIR框架基因%基因分型
殺傷細胞免疫毬蛋白樣受體%流式序列特異性寡覈苷痠探針%序列特異性引物-PCR%KIR框架基因%基因分型
살상세포면역구단백양수체%류식서렬특이성과핵감산탐침%서렬특이성인물-PCR%KIR광가기인%기인분형
killer immunoglobulin-like receptor (KIR)%flow reverse sequence specific oligonucleotide probe (Flow-rSSO)%sequence specific primer-PCR(PCR-SSP)%KIR frame gene%genotyping
目的 探讨常用的序列特异性引物-PCR(PCR-SSP)及流式序列特异性寡核苷酸探针(Flow-rSSO)杂交方法在杀伤细胞免疫球蛋白样受体(KIR)框架基因分型结果中的符合率和准确性.方法 随机选择2011年6月77例深圳造血干细胞捐献者的外周血样,分别用PCR-SSP及Flow-rSSO商品化试剂盒进行KIR框架基因定型,分别通过凝胶电泳图及HLA Fusion 2.0 Research软件分析法,分析结果的一致性.对初检结果不一致的样本,采用同一厂家、不同批号的商品化试剂盒进行复检,并采用美国国立卫生研究院癌症研究所KIR PCR-SSP方法进行检测.结果 77例样本中,75例完全一致,另2例(2.6％) KIR2DL2的结果不一致,PCR-SSP方法对KIR 2DL2的初检、复检检测结果均为阴性,但用Flow-rSSO Lot#004批号试剂盒检测结果为阳性.更换新的Flow-rSSO Lot# 005批号试剂盒复检,结果与PCR-SSP方法的检测结果相一致.结论 为保证KIR基因分型的准确性,开展KIR基因分型的室内、室间质控工作至关重要.
目的 探討常用的序列特異性引物-PCR(PCR-SSP)及流式序列特異性寡覈苷痠探針(Flow-rSSO)雜交方法在殺傷細胞免疫毬蛋白樣受體(KIR)框架基因分型結果中的符閤率和準確性.方法 隨機選擇2011年6月77例深圳造血榦細胞捐獻者的外週血樣,分彆用PCR-SSP及Flow-rSSO商品化試劑盒進行KIR框架基因定型,分彆通過凝膠電泳圖及HLA Fusion 2.0 Research軟件分析法,分析結果的一緻性.對初檢結果不一緻的樣本,採用同一廠傢、不同批號的商品化試劑盒進行複檢,併採用美國國立衛生研究院癌癥研究所KIR PCR-SSP方法進行檢測.結果 77例樣本中,75例完全一緻,另2例(2.6％) KIR2DL2的結果不一緻,PCR-SSP方法對KIR 2DL2的初檢、複檢檢測結果均為陰性,但用Flow-rSSO Lot#004批號試劑盒檢測結果為暘性.更換新的Flow-rSSO Lot# 005批號試劑盒複檢,結果與PCR-SSP方法的檢測結果相一緻.結論 為保證KIR基因分型的準確性,開展KIR基因分型的室內、室間質控工作至關重要.
목적 탐토상용적서렬특이성인물-PCR(PCR-SSP)급류식서렬특이성과핵감산탐침(Flow-rSSO)잡교방법재살상세포면역구단백양수체(KIR)광가기인분형결과중적부합솔화준학성.방법 수궤선택2011년6월77례심수조혈간세포연헌자적외주혈양,분별용PCR-SSP급Flow-rSSO상품화시제합진행KIR광가기인정형,분별통과응효전영도급HLA Fusion 2.0 Research연건분석법,분석결과적일치성.대초검결과불일치적양본,채용동일엄가、불동비호적상품화시제합진행복검,병채용미국국립위생연구원암증연구소KIR PCR-SSP방법진행검측.결과 77례양본중,75례완전일치,령2례(2.6％) KIR2DL2적결과불일치,PCR-SSP방법대KIR 2DL2적초검、복검검측결과균위음성,단용Flow-rSSO Lot#004비호시제합검측결과위양성.경환신적Flow-rSSO Lot# 005비호시제합복검,결과여PCR-SSP방법적검측결과상일치.결론 위보증KIR기인분형적준학성,개전KIR기인분형적실내、실간질공공작지관중요.
Objective To explore the accuracy of killer immunoglobulin-like receptor (KIR) genotyping by using the sequence specific primer-PCR (PCR-SSP)and flow reverse sequence specific oligonucleotide probe(Flow-rSSO)assays.Methods A total of 77 samples from randomly selected stem cell volunteer donors in Shenzhen were subjected to KIR genotyping using the commercial PCR-SSP and Flow-rSSO kits,respectively.The accordance of KIR genotyping results was evaluated and the reason causing the different results was analyzed in this study.Samples with inconclusive and different genotyping results were retested using the same PCR SSP and Flow-rSSO commercial kits with updated lot number and also typed by PCR-SSP method established by National Cancer Institute. Results In the 77 tested samples,the KIR genotypes of 75 samples were in accordance by PCR-SSP and Flow rSSO commercial kit. However,the other two samples showed different results at KIR2DL2 frame gene:the commercial and NC1 PCR-SSP genotyping results indicated the KIR2DL2 were absent,whereas KIR2DL2 were defined present by Flow-rSSO using the initial Lot # 004 commercial kit.We used the updated Flow-rSSO Lot # 005 commercial kit,KIR2DL2 were demonstrated to be absent for the two samples,which is in accordance with results typed by PCR-SSP.Conclusions This study implied that the internal and external quality control would be critical important and,essential in KIR genotyping.
