A new single-step quantitative pathogen detection system: Template-tagging followed by multiplex asymmetric PCR using common primers and CE-SSCP
SCIE
SCOPUS
- Title
- A new single-step quantitative pathogen detection system: Template-tagging followed by multiplex asymmetric PCR using common primers and CE-SSCP
- Authors
- Shin, GW; Cho, YS; Hwang, HS; Oh, MH; Nam, HG; Park, JH; Jung, GY
- Date Issued
- 2009-08
- Publisher
- WILEY-V C H VERLAG GMBH
- Abstract
- Rapid diagnosis of bacterial infection is important for patient management and appropriate therapy during the early phase of bacteria-induced disease. Among the existing techniques for identifying microbial, CE-SSCP combined with 16S ribosomal RNA gene-specific PCR has the benefits of excellent sensitivity, resolution, and reproducibility. However, even though CE-SSCP can separate PCR products with high-resolution, multiplex detection and quantification are complicated by primer-dimer formation and non-specific amplification. Here, we describe a novel technique for multiplex detection and quantification of pathogens by template-tagging followed by multiplex asymmetric PCR and subsequent CE-SSCP. More specifically, we reverse transcribed 16S ribosomal RNAs from seven septicemia-inducing pathogens, tagged the templates with common end sequences, and amplified them using common primers. The resulting amplicons could be successfully separated by CE-SSCP and quantified by comparison to an internal standard. This method yielded results that illustrate the potential of this system for diagnosing infectious disease.
- Keywords
- CE-SSCP; Common primer; Diagnosis; Pathogen; Template-tagging; RIBOSOMAL-RNA GENE; REAL-TIME PCR; DNA MICROARRAY; MICROBIAL COMMUNITY; BACTERIAL PATHOGENS; IDENTIFICATION; ASSAY; AMPLIFICATION; DIAGNOSTICS; SAMPLES
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/26271
- DOI
- 10.1002/ELPS.200900074
- ISSN
- 0173-0835
- Article Type
- Article
- Citation
- ELECTROPHORESIS, vol. 30, no. 15, page. 2728 - 2736, 2009-08
- Files in This Item:
- There are no files associated with this item.
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.