CAJ | 학술논문

为评估定量荧光PCR(Quantitative fluorescent polymerase chain reaction, QF-PCR)技术在快速筛查无精子症因子(Azoospermia factor, AZF)微缺失中的应用,文章对1218例非梗阻性无精子症、少精子症的男性不育患者,采用多重QF-PCR结合毛细管电泳技术,检测Y染色体长臂AZF区9个序列标签位点(Sequence tagged site, STS)以及性染色体短臂的AMEL(Amelogenin)和SRY(Sex-determining region of Y chromosome)位点,辅以常规染色体G显带方法进行核型分析。结果显示,1218例患者中105例可见AZF区微缺失(8.62%),其中AZFc区缺失(67.62%)最常见,其次为AZFb,c区缺失(20.95%)；AZFb区缺失(7.62%)和AZFa区缺失(3.81%)则较少见；另有5例患者为AZFa,b,c区缺失合并AMEL-Y缺失,提示可能缺少Y染色体,经核型分析验证为46,XX(性反转)。105例AZF区微缺失患者的染色体核型分析显示染色体异常16例,其中“Yqh-”12例。根据AMEL-X/AMEL-Y比值,可见1218例患者中86例可能存在性染色体异常,经核型分析验证,68例为性染色体非整倍体。多重QF-PCR 技术,一个反应即能检测样本的多个位点,并可提示性染色体是否存在异常,有助于男性不育患者尽早明确病因,也为后续的检查和治疗提供依据。
위평고정량형광PCR(Quantitative fluorescent polymerase chain reaction, QF-PCR)기술재쾌속사사무정자증인자(Azoospermia factor, AZF)미결실중적응용,문장대1218례비경조성무정자증、소정자증적남성불육환자,채용다중QF-PCR결합모세관전영기술,검측Y염색체장비AZF구9개서렬표첨위점(Sequence tagged site, STS)이급성염색체단비적AMEL(Amelogenin)화SRY(Sex-determining region of Y chromosome)위점,보이상규염색체G현대방법진행핵형분석。결과현시,1218례환자중105례가견AZF구미결실(8.62%),기중AZFc구결실(67.62%)최상견,기차위AZFb,c구결실(20.95%)；AZFb구결실(7.62%)화AZFa구결실(3.81%)칙교소견；령유5례환자위AZFa,b,c구결실합병AMEL-Y결실,제시가능결소Y염색체,경핵형분석험증위46,XX(성반전)。105례AZF구미결실환자적염색체핵형분석현시염색체이상16례,기중“Yqh-”12례。근거AMEL-X/AMEL-Y비치,가견1218례환자중86례가능존재성염색체이상,경핵형분석험증,68례위성염색체비정배체。다중QF-PCR 기술,일개반응즉능검측양본적다개위점,병가제시성염색체시부존재이상,유조우남성불육환자진조명학병인,야위후속적검사화치료제공의거。
To assess the application of quantitative fluorescent polymerase chain reaction (QF-PCR) on rapid screening of azoospermia factor (AZF) microdeletions, 1218 infertile men with non-obstructive azoospermia or oligospermia were de-tected for 9 sequence tagged sites (STSs) in AZF region by multiplex QF-PCR combined with capillary electrophoresis. AMEL (amelogenin) as well as SRY (sex-determining region of Y chromosome) located on short arm of sex chromosome was selected as internal control. Karyotyping was performed on Giemsa-banded metaphase chromosomes of peripheral blood lymphocytes. Of the 1218 patients, 105 (8.62%) were identified as AZF microdeletions. Deletion of AZFc (67.62%) was the most frequent, followed by deletion of AZFb,c (20.95%), AZFb (7.62%) and AZFa (3.81%). Five patients pre-sented with deletions of both AZFa,b,c and AMEL-Y, indicating sex reversal which was confirmed to be 46,XX by karyo-typing. Among the 105 patients with AZF microdeletions, 16 were karyotyped as chromosomal anomalies, most commonly 46,XY,Yqh-(75%, 12/16). In addition, of the total 1218 patients examined, 86 patients showed abnormal AMEL-X/AMEL-Y ratio, suggesting a possibility of sex chromosome anomalies, and 68 of them were verified as sex chromosome aneuploid by karyotyping. Multiplex QF-PCR is capable to detect all markers in one reaction and is also suggestive for sex chromo-some anomalies. It could serve as an effective technique for screening Y-microdeletions, and thus have general application in diagnosis and treatment of male infertility.