Dramatic increase in signal by integration of polymerase chain reaction and hybridization on surface of DNA microarray.
The cumbersome process required for diagnosis by DNA microarray can be simplified by simple extraction of nucleic acid from cells and by integration of liquid-phase polymerase chain reaction (PCR) and hybridization on the surface of a microarray slide. An unexpected benefit was the large (five- to sixfold) increase in detection signal that also is translated into an increase in sensitivity and the confidence level of diagnosis. The large increase in the detection signal appears to be due to participation of PCR primers as well as to extension of the immobilized capture probes during the hybridization process. The reason for the large increase in signal is not clear in view of only one round of DNA synthesis during the hybridization step. The integrated process correctly identified various genotypes of human papillomavirus (HPV) in the infected clinical human cervical specimens with specificity and efficiency. The process described in this article saves labor, time, and cost and should be applicable for automation of diagnosis by DNA microarray.
Kim
Kyung-Tae
KT
Department of Biomedical Science and Technology, Institute of Biomedical Science and Technology, Konkuk University, Seoul 143-701, Republic of Korea.
Na
Chan Hyun
CH
Yun
Yeo Min
YM
Hwang
Tae Sook
TS
Kim
Soo Nyung
SN
Chae
Chi-Bom
CB
D000873:Anthracenes / Q000378:metabolism; D001483:Base Sequence; D002457:Cell Extracts; D045744:Cell Line, Tumor; D002584:Cervix Uteri / Q000473:pathology / Q000821:virology; D004247:DNA / Q000302:isolation & purification; D015342:DNA Probes / Q000378:metabolism; D005260:Female; D015894:Genome, Human / Q000235:genetics; D005838:Genotype; D052162:Human papillomavirus 16 / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D012680:Sensitivity and Specificity; D013499:Surface Properties
D000873:Anthracenes; D002457:Cell Extracts; D015342:DNA Probes; C501244:dendron; D004247:DNA