- Designing Primers from Multiple Sequences Using Matchup Program to Improve Detection of Hepatitis B Virus by Polymerase Chain Reaction
-
So Young Jang , Mi Suk Kim , Min Seok Park , Keon Myung Lee , Hwan Won Chung , Jongsik Chun , Chan Hee Lee
-
J. Microbiol. 2010;48(1):111-116. Published online March 11, 2010
-
DOI: https://doi.org/10.1007/s12275-009-0282-8
-
-
767
View
-
0
Download
-
1
Crossref
-
Abstract
PDF
-
Traditionally primers for PCR detection of viruses have been selected from genomic sequence of single or representative viral strain. However, high mutation rate of viral genomes often results in failure in detecting viruses in clinical and environmental samples. Thus, it seems necessary to consider primers designed from multiple viral sequences in order to improve detection of viral variants. Matchup is a program intended to select universal primers from multiple sequences. We designed using Matchup program primer pairs for HBV detection from 691 full genomic HBV DNA sequences available from NCBI GenBank database. Thousands of primer candidates were initially extracted and these were sequentially filtered down to 5 primer pairs. These primer pairs were tested by PCR using 5 HBV Korean HBsAg(+) patient sera, and eventually one universal primer pair was selected and named MUW (multiple-universal-worldwide). This primer pair, 3 HBV reference primer pairs reported by others and 1 commercial primer pair were compared using 86 HBV
HBsAg(+) sera from Korean and Vietnamese patients. The detection rate for MUW primer pair was 72.1%, much greater than those obtained by reference and commercial primers (32.5 to 40.7%). The superiority of MUW primer pair appeared to be correlated with the conserved sequences of the forward primer binding sites and primer quality score. These results suggest that the universal primers designed by the Matchup program from multiple sequences could be useful in detecting viruses from clinical samples.
-
Citations
Citations to this article as recorded by 
- Development of a new duplex real-time polymerase chain reaction assay for hepatitis B viral DNA detection
Shipeng Sun, Shuang Meng, Rui Zhang, Kuo Zhang, Lunan Wang, Jinming Li Virology Journal.2011;[Epub] CrossRef
- Characterization and Signature Pattern Analysis of Korean Clade HIV-1 Using nef Gene Sequences
-
Chan Seung Park , Dong Hun Lee , Keon Myung Lee , Chan-Hee Lee
-
J. Microbiol. 2008;46(1):88-94.
-
DOI: https://doi.org/10.1007/s12275-007-0156-x
-
-
749
View
-
0
Download
-
3
Crossref
-
Abstract
PDF
-
Phylogenetic studies of the HIV-1 gene sequences isolated from Korean patients have suggested that most of Korean isolates belong to the subtype B strain. This study aims to characterize the Korean clade by molecular phylogenetic analysis using all of the Korean nef gene sequences registered in the NCBI GenBank (N=422), in addition to 41 reference strains and 94 foreign isolates. Through phylogenetic analyses, we verified that most of the Korean isolates belonged to the subtype B, where 78.8% are clustered exclusively of foreign isolates. This cluster has been named the Korean clade subtype B (KCB) in order to distinguish it from other subtype B clusters. Genetic distance analysis suggested that the KCB cluster was more homogeneous and clearly distinctive from the non-Korean clade subtype B (NKCB). Comparison of consensus amino acid sequences from KCB and NKCB revealed that characteristic KCB signature amino acid patterns composed of 11 amino acid residues, whose frequencies in the KCB were significantly higher than in the NKCB. The KCB signature amino acid residues were critical in identifying KCB from NKCB, since substitution of the NKCB sequences with KCB signature amino acids relocated them to the Koran clade, and vice versa.
-
Citations
Citations to this article as recorded by 
- Sequence Length of HIV-1 Subtype B Increases over Time: Analysis of a Cohort of Patients with Hemophilia over 30 Years
Young-Keol Cho, Jung-Eun Kim, Brian T. Foley Viruses.2021; 13(5): 806. CrossRef - Characterization and phylogenetic analysis of Varicella-zoster virus strains isolated from Korean patients
Min Ho Kim, Jeong Seon Jeon, In Kyo Kim, Ji Seon Park, Hosun Park, Ok Sarah Shin, Chan Hee Lee Journal of Microbiology.2017; 55(8): 665. CrossRef - Timing and evolution of the most recent common ancestor of the Korean clade HIV subtype B based on Nef and Vif sequences
Mi-Suk Kim, So-Young Jang, Chan-Seung Park, Keon-Myung Lee, Dong-Hun Lee, Chan-Hee Lee The Journal of Microbiology.2009; 47(1): 85. CrossRef
- Analysis of Substitution Events in HIV-1 vif Gene of the Korean Clade
-
Chan Seung Park , Mi Sook Kim , Hyun Ah Yi , Dong Hun Lee , Keon Myung Lee , Chan Hee Lee
-
J. Microbiol. 2007;45(1):75-78.
-
DOI: https://doi.org/2487 [pii]
-
-
Abstract
PDF
-
Nucleotide and amino acid substitution pattern in vif gene of the Korean clade of HIV-1 isolated from Koreans were analyzed using consensus sequences. At nucleotide level, transition/transversion substitution ratio was 1.88, and nonsynonymous/synonymous substitution ratio was 2.67, suggesting a divergent
<br>evolution in the Korean clade. At amino acid level, there were 17 substitutions and G→E substitution at position 37 may be responsible for change in predicted secondary structure.
- Molecular Phylogenetic Analysis of HIV-1 vif Gene from Korean Isolates
-
Chan Seung Park , Mi Sook Kim , Sung Duk Lee , Sung Soo Kim , Keon Myung Lee , Chan-Hee Lee
-
J. Microbiol. 2006;44(6):655-659.
-
DOI: https://doi.org/2461 [pii]
-
-
Abstract
PDF
-
Phylogenetic studies of nef, pol, and env gene sequences of HIV-1 isolated from Koreans suggested the presence of a Korean clade in which Korean sequences are clustered to the exclusion of foreign sequences. We attempted to identify and characterize the Korean clade using all vif gene sequences isolated from Koreans registered in the NCBI GenBank database (n = 233). Most (77%) of the Korean isolates belonged to the Korean clade as a large subcluster in subtype B, designated the Korean clade subtype B (KCB). KCB sequences were relatively homogenous compared to Korean subtype B sequences that did not belong to the KCB (non-Korean clade subtype B; NKCB). Comparison of amino acid frequencies of KCB and NKCB sequences revealed several positions where the amino acid frequencies were significantly different. These amino acid residues were critical in separating KCB from NKCB or from foreign sequences, since substitution of these amino acids in KCB with the NKCB amino acids relocated the KCB sequences to NKCB, and vice versa. Further analyses of KCB will help us to understand the origin and evolutionary history of KCB.
|