Simple and versatile molecular method of copy-number measurement using cloned competitors.
Variations and alterations of copy numbers (CNVs and CNAs) carry disease susceptibility and drug responsiveness implications. Although there are many molecular methods to measure copy numbers, sensitivity, reproducibility, cost, and time issues remain. In the present study, we were able to solve those problems utilizing our modified real competitive PCR method with cloned competitors (mrcPCR). First, the mrcPCR for ERBB2 copy number was established, and the results were comparable to current standard methods but with a shorter assay time and a lower cost. Second, the mrcPCR assays for 24 drug-target genes were established, and the results in a panel of NCI-60 cells were comparable to those from real-time PCR and microarray. Third, the mrcPCR results for FCGR3A and the FCGR3B CNVs were comparable to those by the paralog ratio test (PRT), but without PRT's limitations. These results suggest that mrcPCR is comparable to the currently available standard or the most sensitive methods. In addition, mrcPCR would be invaluable for measurement of CNVs in genes with variants of similar structures, because combination of the other methods is not necessary, along with its other advantages such as short assay time, small sample amount requirement, and applicability to all sequences and genes.
Research Institute, National Cancer Center, Ilsandong-gu, Goyang, Korea.
Hyun-Kyoung
HK
Kim
Hai-Li
HL
Hwang
Seong-Yeol
SY
Park
Kwang Man
KM
Lee
Won Cheol
WC
Park
Han-Seong
HS
Kim
Tae-Hyun
TH
Um
Young Jun
YJ
Hong
Jin Kyung
JK
Lee
Sun-Young
SY
Joo
Ju-Young
JY
Seoh
Yeong-Wook
YW
Song
Soo-Youl
SY
Kim
Yong-Nyun
YN
Kim
Kyeong-Man
KM
Hong
D001943:Breast Neoplasms / Q000235:genetics; D045744:Cell Line, Tumor; D016000:Cluster Analysis; D056915:DNA Copy Number Variations; D005260:Female; D058851:GPI-Linked Proteins / Q000235:genetics; D005784:Gene Amplification; D018628:Gene Dosage; D020869:Gene Expression Profiling; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics; D017452:Receptors, IgG / Q000235:genetics; D015203:Reproducibility of Results
D058851:GPI-Linked Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases; D017452:Receptors, IgG; C496124:FCGR3A protein, human; C428200:FCGR3B protein, human