sRapid determination of sex chromosome abnormality using double relative quantitative SD-qPCR

doi:10.3969/j.issn.1673-8640.2022.06.012

Abstract

Abstract:

Objective To establish a single tube fluorescent double relative segmental duplication-quantitative polymerase chain reaction（SD-qPCR） for determining the number of chromosome X and Y simultaneously. Methods The established SD-qPCR was used for the determination of 197 amniotic fluid samples. The numbers of chromosome X and Y were determined by the relative quantitative 2^-ΔΔCt value between segmental duplications. The results are compared with chromosome karyotype analysis. Results Two pairs of segmental duplication molecular markers were screened and located on chromosome 16 and chromosome X,chromosome 1 and chromosome Y,respectively. Among the 197 samples,there were 185 samples with normal sex chromosome,5 cases of 45,X,3 cases of 47,XXX,2 cases of 47,XXY and 2 cases of 47,XYY. The results of SD-qPCR were consistent with those of chromosome karyotype analysis. Conclusions The established SD-qPCR can determine the numbers of chromosome X and chromosome Y in a single tube simultaneously. It has the advantages of multiple loci and stability,and it can be widely used in clinic.

Key words: Prenatal diagnosis, Aneuploidy, Real-time fluorescent quantitative polymerase chain reaction, Sex chromosome

CLC Number:

R446.1

SUN Lei, LONG Ju, FAN Zuqian, WU Suping, LIN Guixian, LING Xiuming. sRapid determination of sex chromosome abnormality using double relative quantitative SD-qPCR[J]. Laboratory Medicine, 2022, 37(6): 557-560.

Figures/Tables 3

References 15

[1]	VISOOTSAK J, ROSNER B, DYKENS E, et al. Behavioral phenotype of sex chromosome aneuploidies:48,XXYY,48,XXXY,and 49,XXXXY[J]. Am J Med Genet A, 2007, 143A(11):1198-1203.
[2]	吴佩芳, 张卫华, 唐萍, 等. 嘉兴地区无创产前检测性染色体非整倍体病例的妊娠选择及妊娠结局分析[J]. 中国优生与遗传杂志, 2020, 28(7):820-822.
[3]	LINDEN M G, BENDER B G, ROBINSON A. Intrauterine diagnosis of sex chromosome aneuploidy[J]. Obstet Gynecol, 1996, 87(3):468-475.
[4]	QIAO J, YUAN J, HU W, et al. Combined diagnosis of QF-PCR and CNV-Seq in fetal chromosomal abnormalities:a new perspective on prenatal diagnosis[J]. J Clin Lab Anal, 2022, 36(4):e24311.
[5]	WHALEY R D, DOUGHERTY R E, CHENG L, et al. fluorescence in situ hybridization for the X and Y chromosome centromeres helps differentiate between gestational and nongestational choriocarcinoma in clinically ambiguous cases[J]. Arch Pathol Lab Med, 2020, 144(7):863-868.
[6]	DAI X, SHI F, CHEUNG C K Y, et al. Abnormal Y chromosome detection in infertile males using multiplex ligation-dependent probe amplification[J]. Andrologia, 2022, 54(2):e14316.
[7]	陈杏园, 罗世强, 王秋华, 等. 早孕期流产胚胎组织MLPA检测结果及与年龄和孕周的关系分析[J]. 分子诊断与治疗杂志, 2020, 12(10):1289-1293.
[8]	DUDAREWICZ L, HOLZGREVE W, JEZIOROWSKA A, et al. Molecular methods for rapid detection of aneuploidy[J]. J Appl Genet, 2005, 46(2):207-215.
[9]	HELMY S M, ISMAIL S, BASSIOUNI R, et al. Sensitivity of DCSR3/GAPDH ratio using quantitative real-time PCR in the rapid prenatal diagnosis for down syndrome[J]. Fetal Diagn Ther, 2009, 25(2):220-223.
[10]	SUN L, FAN Z, WENG X, et al. Rapid detection of Down's syndrome using quantitative real-time PCR(qPCR) targeting segmental duplications on chromosomes 21 and 11[J]. Gene, 2014, 552(2):272-276.
[11]	LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method[J]. Methods, 2001, 25(4):402-408.
[12]	CHANDNA R, AUGUSTINE R, BISHT N C. Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR[J]. PLoS One, 2012, 7(5):e36918.
[13]	史宏志, 达布西力特, 高宏凯, 等. 长链非编码RNA p2819在胃癌组织中的差异表达及其意义[J]. 肿瘤综合治疗电子杂志, 2021, 7(1):47-50.
[14]	KONG X, LI L, SUN L, et al. Rapid diagnosis of aneuploidy using segmental duplication quantitative fluorescent PCR[J]. PLoS One, 2014, 9(3):e88932.
[15]	ZIMMERMANN B, HOLZGREVE W, WENZEL F, et al. Novel real-time quantitative PCR test for trisomy 21[J]. Clin Chem, 2002, 48(2):362-363.