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Fluorotyping of HLA-DRB by sequence-specific priming and fluorogenic probing by M. Albis-Camps; R. Blasczyk is a Immunology and Microbiology article available to read on EtoBox.

What is Fluorotyping of HLA-DRB by sequence-specific priming and fluorogenic probing about?

## Abstract: Similar to our recently described HLA‐A and ‐C fluorotyping strategies, the aim of this study was to develop a sequence‐specific primed polymerase chain reaction (PCR‐SSP)‐based fluorotyping method for HLA‐DRB. Applying the fluorogenic 5′ nuclease assay, it is possible to increase the sample throughput rate by abolishing all labor‐intensive post‐amplification steps. Additionally, problems related to contamination are eliminated. The method relies on the 5′–3′ exonuclease activity of the Taq‐DNA Polymerase which cleaves a target‐specific and individually labelled fluorogenic probe during successful PCR. Different labelled probes specific for different targets can be applied in a single PCR, allowing independent detection of the specific HLA and the internal control product. The probe used to detect the HLA‐DRB specific amplicons was labeled at its 5′ end with FAM as the reporter and further 3′ with TAMRA as the quencher. The probe hybridized within the 2nd exon to a conserved region which was covered by all primer mixes. In case of amplification, the cleavage of the fluorogenic probe led to an interruption of the TAMRA‐mediated quenching effect and generated a significa

Who reads Fluorotyping of HLA-DRB by sequence-specific priming and fluorogenic probing?

Author

M. Albis-Camps; R. Blasczyk

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); Blackwell Publishing Inc.; Wiley (ISSN 0001-2815)

Published

1999

Language

EN

Field

Immunology and Microbiology (Life Sciences)