[Clinical application of SYBR Green I real-time fluorescence quantitative PCR for diagnosis of common chromosome aneuploidy].
OBJECTIVE
To investigate the value of real-time fluorescence quantitative PCR in the diagnosis of chromosome anepuploidy.


METHODS
ABCC4 gene on chromosome 13, TYMS gene on chromosome 18, DSCR3 gene on chromosome 21, HPRT2 gene on chromosome X, and SRY gene on Y chromosome were used as the target genes, with GAPDH gene on chromosome 12 as the control gene. Using double-standard curve fluorescent relative quantitative PCR method with SYBR Green as the fluorescent dye, the gene expression levels were detected and the results were compared with those of karyotype analysis.


RESULTS
The ratio of the target gene on chromosome 13 to the control gene showed a significant difference between the normal karyotype group (0.90 - or + 0.31) and trisome group (1.39 - or + 0.12, P=0.003), and the genes on chromosome 18 (1.07 - or + 0.44 vs 1.66 - or + 0.12, P=0.000) and chromosome 21 (0.84 - or + 0.27 vs 1.73 - or + 0.54, P=0.000) showed similar results. The expression of the genes on the X chromosome showed no significant difference between 45, X group and 46,XY group (0.62 - or + 0.12 vs 0.63 - or + 0.25, P=0.965), nor between 46, XX group and 47,XXY group (1.32 - or + 0.37 vs 1.20 - or + 0.35, P=0.326), while a significant difference was noted between the single copy X (including 45,X and 46,XY) and two copies X (46,XX and 47,XXY) (0.63 - or + 0.23 vs 1.26 - or + 0.36, P=0.000). The expression of the target gene on the Y chromosome was not detected in normal females (46,XX), and a significant difference in the expression was found between normal male group (46,XY) and 47,XYY group (1.57 - or + 0.54 vs 3.08 - or + 0.15, P=0.003).


CONCLUSIONS
SYBR Green I real-time fluorescence quantitative PCR can be used for the purpose of rapid diagnosis of chromosome aneuploidy.
Liu
Li-juan
LJ
Basic Medical Experiment Teaching Center, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, China. liulijuan.1983@163.com
Xiong
Li
L
Liu
Jie
J
Deng
Kang
K
Liu
Si-ping
SP
Wu
Rui-feng
RF
Jia
Bei
B
Song
Lan-lin
LL
Zhong
Mei
M
Zeng
Rong
R
D000782:Aneuploidy; D052160:Benzothiazoles; D025063:Chromosome Disorders / Q000175:diagnosis; D002882:Chromosomes, Human, Pair 13 / Q000235:genetics; D002887:Chromosomes, Human, Pair 18 / Q000235:genetics; D002891:Chromosomes, Human, Pair 21 / Q000235:genetics*; D003959:Diamines; D005260:Female; D005453:Fluorescence; D006801:Humans; D008297:Male; D009930:Organic Chemicals; D011804:Quinolines; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014314:Trisomy / Q000175:diagnosis*
D052160:Benzothiazoles; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I