ELECTROCHEMICAL DNA BIOSENSORS phần 5 pptx

HSV Type I infects the nervoussystem, however HSV Type II infects the genital system.. In routine analysis; the discrimination between Type I and II is done by sequence detection syste

Herpes Simplex Virus;

Type I PCR Product

5’TCAACTTCGACTGGCCCTTC T T G CTG G CCAAGCTGACG

GA C AT T TACAAGGTCCC C CT G GAGACGGGTACGG C CGCA TGAACGGCCGGGG C GTGTT T CGCGTGTGGGACATAGGCC AGAGCCACTTCCAGAAGCGCAGCAAGATAAAGGTGAACG

GCATGGTGA G CATCGACATGTACGG 3’

Type II PCR Product

5’TCAACTTCGACTGGCCCTTC G T C CTG A CCAAGCTGACG

GA G AT C TACAAGGTCCC G CT C GAGACGGGTACGG G CGCA TGAACGGCCGGGG T GTGTT C CGCGTGTGGGACATAGGCC AGAGCCACTTCCAGAAGCGCAGCAAGATAAAGGTGAACG

GCATGGTGA A CATCGACATGTACGG 3’

HSV Type I infects the nervous

system, however HSV Type II infects the

genital system.

In routine analysis; the discrimination between Type I

and II is done by sequence

detection system.

Variations of HSV Type 1 and Type 2 discriminations and probes selection :

HSV Type 1 (between 64208 and 64386 bases sequences)

5’ ATC AAC TTC GAC TGG CCC TTC * TTG CTG GCC AAG CTG ACG GAC A * TT TAC AAG GTC CCC CTG GAC GGG TAC GGC CGC ATG AAC GGC CGG GGC GTG TTT CGC GTG TGG GAC ATA GGC CAG AGC CAC TTC CAG AAG CGC AGC AAG ATA AAG GTG AAC GGC ATG GTG AGC ATC

GAC ATG TAC GG 3’

HSV Type 2 (between 64669 and 64847 bases sequences)

5’ ATC AAC TTC GAC TGG CCC TTC *GTC CTG ACC AAG CTG ACG GAG A * TC TAC AAG GTC CCG CTC GAC GGG TAC GGG CGC ATG AAC GGC CGG GGT GTG TTC CGC GTG TGG GAC ATC GGC CAG AGC CAC TTC CAG AAG CGC AGC AAG ATA AAG GTG AAC GGC ATG GTG AAC

ATC GAC ATG TAC GG 3’

Real Samples-2

Label-Free Electrochemical Hybridization Genosensor for the Detection of Hepatitis

B Virus Genotype on the Development of

Lamivudine Resistance

-Lamivudine is a pyrimidine nucleoside analogue that

inhibits viral replication by blocking viral reverse transcriptase

-Continuous lamivudine therapy may lead to selection of

resistant strains

Anal Chem.2005, 77,4908-4917

z Hepatitis B virus (HBV) causes acute and chronic hepatitis, chirrosis and

hepatocelular carcinoma.

z Hepatitis B virus infection is the ninth

mainly reason of death on the world

According to the World Health Organization (WHO) more than 400 million people are chronically infected

by Hepatitis B virus (HBV) worldwide

Long-term treatment is often needed and that is

associated with many problems such as,

z Lamivudine-resistance is associated with nucleotide

substitutions that induce amino acid changes in codon 204 of the polymerase gene

instead of methionine in the tyrosine(Y), methionine(M),

aspartate(D) and aspartate(D) motif (YMDD motif)

general described mutations

PCR MAP of HBV

catcctgctg ctatgcctca tcttcttgtt ggttcttctg gattatcaag

gtatgttgcc cgtttgtcct ctaattccag gatcaacaac aaccagtacg

ggaccatgca aaacctgcac gactcctgct caaggcaact ctatgtttcc

ctcatgttgc tgtacaaaac ctacggatgg aaattgcacc tgtattccca

tcccatcgtc ctgggctttc gcaaaatacc tatgggagtg ggcctcagtc cgtttctctt ggctcagttt actagtgcca tttgttcagt ggttcgtagg

ggccaagact gtacagcatc gtgagtccct ttataccgct

z YMDD mutants are caused by a point mutation from A to

G at the 743nd position (YVDD) and from G to T at the 745rd position (YIDD) of the human genome