PMID- 17691029
OWN - NLM
STAT- MEDLINE
DCOM- 20071002
LR  - 20211203
IS  - 0882-0139 (Print)
IS  - 0882-0139 (Linking)
VI  - 36
IP  - 4
DP  - 2007
TI  - Rapid HLA-DR fluorotyping based on melting curve analysis.
PG  - 507-21
AB  - Real-time polymerase chain reaction (PCR) has been used in the study of human 
      leukocyte antigen (HLA) genotyping as a potential alternative for routinely used 
      molecular methods such as PCR-sequence specific primers (PCR-SSP) and 
      PCR-sequence specific oligonucleotide probes (PCR-SSO). Combined with fluorescent 
      dye like SYBR GREEN I, it has more advantages such as low cost and consistent 
      background. The aim of this study was to optimize the fluorescent dye-based 
      method and introduce it into the fluorotyping for HLA-DR lotus. 24 pairs of 
      allele-specific primers and 1 pair of internal control were optimized to 
      discriminate HLA-DRB1, -DRB3/B4/B5 alleles. Additionally, conditions of real-time 
      PCR amplifying and melting curve recording had been improved for convenient and 
      clear readout. Forty-two clinical samples previously typed by conventional 
      PCR-SSP or sequence based typing (SBT) were tested and all got identical results. 
      With this technique, 15 DNA samples can be assayed in parallel within 2 hours on 
      the Real-time PCR instrument. These data strongly suggest a rapid HLA-DR 
      fluorotyping method based on melting curve analysis, which could be a more 
      economic and automatic alternative for clinical HLA-DR typing.
FAU - Liu, Yang
AU  - Liu Y
AD  - Laboratory of Molecular Immunology, State Key Laboratory of Genetic Engineering, 
      School of Life Sciences, Fudan University, Shanghai, China.
FAU - Zhu, Qiyun
AU  - Zhu Q
FAU - Zhu, Naishuo
AU  - Zhu N
LA  - eng
PT  - Journal Article
PL  - England
TA  - Immunol Invest
JT  - Immunological investigations
JID - 8504629
RN  - 0 (Benzothiazoles)
RN  - 0 (Diamines)
RN  - 0 (HLA-DR Antigens)
RN  - 0 (Organic Chemicals)
RN  - 0 (Quinolines)
RN  - 163795-75-3 (SYBR Green I)
SB  - IM
MH  - Alleles
MH  - Benzothiazoles
MH  - Diamines
MH  - HLA-DR Antigens/*immunology
MH  - Histocompatibility Testing
MH  - Humans
MH  - Organic Chemicals
MH  - Polymerase Chain Reaction/*methods
MH  - Quinolines
MH  - Sensitivity and Specificity
MH  - Transition Temperature
EDAT- 2007/08/11 09:00
MHDA- 2007/10/03 09:00
CRDT- 2007/08/11 09:00
PHST- 2007/08/11 09:00 [pubmed]
PHST- 2007/10/03 09:00 [medline]
PHST- 2007/08/11 09:00 [entrez]
AID - 781212853 [pii]
AID - 10.1080/08820130701366425 [doi]
PST - ppublish
SO  - Immunol Invest. 2007;36(4):507-21. doi: 10.1080/08820130701366425.