山西职工医学院学报
山西職工醫學院學報
산서직공의학원학보
JOURNAL OF SHANXI MEDICAL COLLEGE FOR CONTINUING EDUCATION
2014年
4期
8-13
,共6页
HBV%BCP区双突变%前C区突变%MAMA-PCR
HBV%BCP區雙突變%前C區突變%MAMA-PCR
HBV%BCP구쌍돌변%전C구돌변%MAMA-PCR
HBV%BCP double mutation%precore mutation%MAMA-PCR
目的：利用错配扩增和荧光PCR原理，建立一种针对HBV BCP区1762/1764和前C区1896突变位点的新型检测方法---错配扩增突变分析PCR法（ MAMA-PCR），并对该方法进行临床评价。方法：a）MAMA-PCR突变检测方法性能评估：以野生和突变性阳性参考品作为模板检测3次，分析该方法稳定性；将突变型和野生型质控品梯度比例混合，分析该方法检测混合感染的检测效率。b）测序法、商品化试剂盒与本方法同时检测样本并对MAMA-PCR方法做出评价。结果：MAMA-PCR 3次检测重复性良好，CV值均小于5%，且可稳定检出含1%突变含量的混合样本。132例HBV样本中，1762/1764双突变位点测序法检出67例，ARMS-PCR试剂盒检出69例，MAMA-PCR检出69例，突变检出率分别为50.8%、52.3%和52.3%；1896突变测序法检出39例，ARMS-PCR试剂盒检出41例，MAMA-PCR检出42例，突变检出率分别为29.5%、31.1%和31.8%。结论：MAMA-PCR 检测方法性能稳定，突变检出率高于测序法，与商用试剂盒基本一致，可用于临床乙肝患者 BCP区1762/1764双突变和前C区1896突变位点的快速检测。
目的：利用錯配擴增和熒光PCR原理，建立一種針對HBV BCP區1762/1764和前C區1896突變位點的新型檢測方法---錯配擴增突變分析PCR法（ MAMA-PCR），併對該方法進行臨床評價。方法：a）MAMA-PCR突變檢測方法性能評估：以野生和突變性暘性參攷品作為模闆檢測3次，分析該方法穩定性；將突變型和野生型質控品梯度比例混閤，分析該方法檢測混閤感染的檢測效率。b）測序法、商品化試劑盒與本方法同時檢測樣本併對MAMA-PCR方法做齣評價。結果：MAMA-PCR 3次檢測重複性良好，CV值均小于5%，且可穩定檢齣含1%突變含量的混閤樣本。132例HBV樣本中，1762/1764雙突變位點測序法檢齣67例，ARMS-PCR試劑盒檢齣69例，MAMA-PCR檢齣69例，突變檢齣率分彆為50.8%、52.3%和52.3%；1896突變測序法檢齣39例，ARMS-PCR試劑盒檢齣41例，MAMA-PCR檢齣42例，突變檢齣率分彆為29.5%、31.1%和31.8%。結論：MAMA-PCR 檢測方法性能穩定，突變檢齣率高于測序法，與商用試劑盒基本一緻，可用于臨床乙肝患者 BCP區1762/1764雙突變和前C區1896突變位點的快速檢測。
목적：이용착배확증화형광PCR원리，건립일충침대HBV BCP구1762/1764화전C구1896돌변위점적신형검측방법---착배확증돌변분석PCR법（ MAMA-PCR），병대해방법진행림상평개。방법：a）MAMA-PCR돌변검측방법성능평고：이야생화돌변성양성삼고품작위모판검측3차，분석해방법은정성；장돌변형화야생형질공품제도비례혼합，분석해방법검측혼합감염적검측효솔。b）측서법、상품화시제합여본방법동시검측양본병대MAMA-PCR방법주출평개。결과：MAMA-PCR 3차검측중복성량호，CV치균소우5%，차가은정검출함1%돌변함량적혼합양본。132례HBV양본중，1762/1764쌍돌변위점측서법검출67례，ARMS-PCR시제합검출69례，MAMA-PCR검출69례，돌변검출솔분별위50.8%、52.3%화52.3%；1896돌변측서법검출39례，ARMS-PCR시제합검출41례，MAMA-PCR검출42례，돌변검출솔분별위29.5%、31.1%화31.8%。결론：MAMA-PCR 검측방법성능은정，돌변검출솔고우측서법，여상용시제합기본일치，가용우림상을간환자 BCP구1762/1764쌍돌변화전C구1896돌변위점적쾌속검측。
Objective:To establish a new method( MAMA-PCR)for the detection of mutations at 1762/1764 and 1896 in the BCP region and precore region of HBV using mismatch amplification and Fluorescent PCR and to evaluate this method with clinical samples. Method:a)Evaluate the parameters of the MAMA-PCR mutation detection method:to analyze the stability of reagents by reactions with three repeats using the wild-type and mutant reference as a template;to test the detection efficiency of the HBV hybrid infection using mutant and wild-type references with different mixing ratio. b)Compare the MAMA-PCR method with sequencing and commercialized mutation detection kits and evaluate the MAMA-PCR method. Results:Three tests of MAMA-PCR had good repeatability with a CV value of less than 5% and HBV infection samples which contained 1% mutation could be stably detected. In 132 HBV samples,sequencing meth-od identified 67 with 1762/1764 mutations,mutation detection kit identified 69,and MAMA-PCR identified 69. The detection rates were 50. 8%,52. 3%,and 52. 3%,respectively. Sequencing method identified 39 samples with 1896 mutation,mutation detection kit identified 41,and MAMA-PCR identified 42. The detection rates were 29. 5%, 31. 1%,and 31. 8%,respectively. Conclusions:The MAMA-PCR method is very stable. The mutation detection rate of MAMA-PCR is almost the same as commercialized kits,and is higher than sequencing. This new technology( MAMA-PCR)can be used in the fast detection of HBV BCP 1762/1764 double mutations and precore 1896 mutation in clinical samples.
