中华检验医学杂志
中華檢驗醫學雜誌
중화검험의학잡지
CHINESE JOURNAL OF LABORATORY MEDICINE
2010年
2期
111-114
,共4页
叶峻杰%阎慧%李宗芳%郭海%王跃力%李江川%赵树华
葉峻傑%閻慧%李宗芳%郭海%王躍力%李江川%趙樹華
협준걸%염혜%리종방%곽해%왕약력%리강천%조수화
不育,男(雄)性%Y染色体%染色体缺失%聚合酶链反应
不育,男(雄)性%Y染色體%染色體缺失%聚閤酶鏈反應
불육,남(웅)성%Y염색체%염색체결실%취합매련반응
Infertility,male%Y chromosome%Chromosome deletion%Polymerase chain reaction
目的 评价改良多重PcR筛查男性非梗阻性无精子症和严重少精子症患者Y染色体AZF缺失类型的价值.方法 在Y染色体STS常规引物序列的5'端连接一段非人类同源序列的寡核苷酸链,合成嵌合引物对.根据非人类同源序列的寡核苷酸链设计通用引物对,与嵌合引物对在同一反应体系中对人类基因组DNA进行多重PCR扩增,并用以评价筛查262例非梗阻性无精子症和严重少精子症患者中AZF a、b、c区域的微缺失状况.结果 用改良多重PCR筛查262例非梗阻性无精子症和严重少精子症患者,发现AZF微缺失33例(12.60%),其中AZF c缺失27例,AZF b+c缺失6例,与EMQN推荐方法检测的结果相比,缺失阳性符合率为100%(33/33),且未出现假阳性.改良多重PCR与EMQN推荐方法检测Y染色体多个STS的电泳结果显示,2种方法得到的sY84、sY86、sY127、sY134、sY254、sY255、SRY位点扩增产物的均一性好,扩增产物条带清晰.结论 改良多重PCR能较好地筛查出男性非梗阻性无精子症和严重少精子症患者Y染色体AZF缺失类型.
目的 評價改良多重PcR篩查男性非梗阻性無精子癥和嚴重少精子癥患者Y染色體AZF缺失類型的價值.方法 在Y染色體STS常規引物序列的5'耑連接一段非人類同源序列的寡覈苷痠鏈,閤成嵌閤引物對.根據非人類同源序列的寡覈苷痠鏈設計通用引物對,與嵌閤引物對在同一反應體繫中對人類基因組DNA進行多重PCR擴增,併用以評價篩查262例非梗阻性無精子癥和嚴重少精子癥患者中AZF a、b、c區域的微缺失狀況.結果 用改良多重PCR篩查262例非梗阻性無精子癥和嚴重少精子癥患者,髮現AZF微缺失33例(12.60%),其中AZF c缺失27例,AZF b+c缺失6例,與EMQN推薦方法檢測的結果相比,缺失暘性符閤率為100%(33/33),且未齣現假暘性.改良多重PCR與EMQN推薦方法檢測Y染色體多箇STS的電泳結果顯示,2種方法得到的sY84、sY86、sY127、sY134、sY254、sY255、SRY位點擴增產物的均一性好,擴增產物條帶清晰.結論 改良多重PCR能較好地篩查齣男性非梗阻性無精子癥和嚴重少精子癥患者Y染色體AZF缺失類型.
목적 평개개량다중PcR사사남성비경조성무정자증화엄중소정자증환자Y염색체AZF결실류형적개치.방법 재Y염색체STS상규인물서렬적5'단련접일단비인류동원서렬적과핵감산련,합성감합인물대.근거비인류동원서렬적과핵감산련설계통용인물대,여감합인물대재동일반응체계중대인류기인조DNA진행다중PCR확증,병용이평개사사262례비경조성무정자증화엄중소정자증환자중AZF a、b、c구역적미결실상황.결과 용개량다중PCR사사262례비경조성무정자증화엄중소정자증환자,발현AZF미결실33례(12.60%),기중AZF c결실27례,AZF b+c결실6례,여EMQN추천방법검측적결과상비,결실양성부합솔위100%(33/33),차미출현가양성.개량다중PCR여EMQN추천방법검측Y염색체다개STS적전영결과현시,2충방법득도적sY84、sY86、sY127、sY134、sY254、sY255、SRY위점확증산물적균일성호,확증산물조대청석.결론 개량다중PCR능교호지사사출남성비경조성무정자증화엄중소정자증환자Y염색체AZF결실류형.
Objective To establish an universal primer-multiplex PCR system for diagnosis of Y chromosome AZF region microdeletions in 262 patients with non-obstructive azoospermic and severe oligozoospermic male infertility. Methods In each panel of multiplex PCR, YUP and YCP containing a fragment of non-human DNA sequence at their 5' ends were designed. The universal primers and chimiric primers were employed for the amplification at the same multiplex PCR system to screen for the Y chromosome AZF region ( a, b and c) microdeletions in 262 non-obstructive azoospermic and severe oligozoospermic male infertility patients. Results Thirty-three out of 262 patients (12. 60% ) were detected with Y chromosome AZF microdeletions. Among them, 27 cases were AZF c microdeletions and 6 ones were AZF b + c microdeletions. These results were in agreement with the results from EMQN method. There was no false-positivity. The gel electrophoresis for detection of multiple STS from both methods showed that the sY84,sY86, sY127, sY134, sY254, sY255, SRY bands were homogeneous and clear with similar brightness. Conclusion The modified multiplex PCR is suitable for screening of Y chromosome AZF microdeletions in non-obstructive azoospermic and severe digozoospermic male infertility patients.
