Biology Reference
In-Depth Information
[51]. A PCR sequence was used related to Hepatitis B (HBV) virus
genome. Optimization of hybridization detection was performed
with 17-mer short oligonucleotides. Carbon paste electrodes (CPE)
and hanging mercury drop electrodes (HMDE) were used as
sensor surface, CV and DPV transduction of MDB accumulated
after hybridization between 23-mer capture probe and HBV-PCR
amplicon was monitored for the detection. By using MDB indicator,
Herpes simplex (HSV) virus genome detection and discrimination
of HSV type I and type II viruses were performed in PCR amplicon
[52]. HSV type I and type II have very similar pathogenesis
mechanisms and have a homogeneous genome sequence. Two
types of PCR products related to type I and II which had 12
base differences in 179 base long amplicon were used as target
genomes. 22-mer capture probes related to type I and II had four
basedifferencesbetweeneachother,wereattachedontodisposable
graphite electrode surfaces and hybridization occurred with both
types of PCR amplicons. The detection and the discrimination of
genotypingwereaccomplishedbyDPVtransductionofaccumulated
MDB. Consequently four base differences were detected by using
long PCR amplicon devoted to clinicalanalysis.
One base mismatch detections in real samples based on MDB
indicator were also performed in our following researches. In 2007,
wedevelopedagenosensorfordetectionoftoll-likereceptor2(TLR
-2) gene polymorphisms [53]. In this study, one base mismatch
detectionwasperformedina267baselongPCRproduct.Twotypes
of capture probes were used representing wild-type and mutant-
type genomes. DPV reduction signals of MDB were monitored after
hybridization with denatured amplicon at PGE surfaces. Heterozy-
gous and homozygous discrimination was also performed by using
twotypesofcaptureprobes.Biosensorselectivitywasachievedwith
HBV non-complementary (NC) amplicon. Consequently an allele
specific genosensor was developed for SNP detection in this study.
AnotherpolymorphismdetectionrelatedtoApaIvitaminDreceptor
gene was also performed in 2010 [54]. DPV signals of accumulated
MDB indicated hybridization and mismatch detection in 247-mer
PCR sequence.
Some hybridization indicators have chemical a
nity to DNA
bases. Our group used another dye molecule, methylene blue
