PMID- 29541460
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20201001
IS  - 2049-9434 (Print)
IS  - 2049-9442 (Electronic)
IS  - 2049-9434 (Linking)
VI  - 8
IP  - 4
DP  - 2018 Apr
TI  - Optimization of an in-house PCR method for the detection of HLA-B*27 alleles.
PG  - 385-390
LID - 10.3892/br.2018.1055 [doi]
AB  - The association of spondyloarthropathies with different alleles of human 
      leukocyte antigen (HLA) B*27 is well established. Different subtypes of HLA-B*27 
      may be linked with different ethnic groups, distinct clinical manifestations, 
      specific age of onset and different prognoses. Polymerase chain reaction with 
      sequence specific primers (PCR-SSP) is the most frequently adapted molecular 
      method used for the recognition of HLA-B*27-specific DNA sequences. The aim of 
      the present study was to standarise an in-house protocol of PCR-SSP for HLA-B*27 
      allele detection for use in the Armed Forces Institute of Pathology (AFIP), 
      Pakistan, with consideration of its cost effectiveness. A total of 49 individual 
      samples were included, comprising 10 transplant samples determined to be 
      HLA-B*27-negative by PCR-SSP and 39 HLA-B*27-positive samples determined by flow 
      cytometry, obtained from patients who were symptomatic and referred for HLA-B*27 
      testing. By altering each variable individually, an in-house PCR-SSP protocol was 
      optimized to amplify common HLA-B*27 alleles (2701-2721, 2723-2730). To 
      discriminate B*27 from all other HLA-B alleles, a low-resolution HLA-B typing set 
      with a 96 PCR-SSP primer mixture was used in conjunction. Among the 39 
      HLA-B*27-positive specimens, 31 (79%) were detected as positive by PCR-SSP, with 
      the remaining samples failing due to a sub-optimized protocol and/or low DNA 
      concentration. Additionally, there was complete concordance between flow 
      cytometry and in-house PCR, and the sensitivity and specificity of the PCR-SSP 
      were determined to be 100%. In conclusion, in-house SSP-PCR is, standard method 
      for the detection of HLA-B*27 alleles. The determination of associations between 
      specific HLA-B*27 alleles and AS may aid to identify individuals at higher risk 
      of developing the disease. Furthermore, the identification of individuals at risk 
      may aid to adapt preventive strategies.
FAU - Afshan, Noor
AU  - Afshan N
AD  - Department of Immunology, Armed Forces Institute of Pathology, 46000 Rawalpindi, 
      Pakistan.
FAU - Bashir, Mukarram
AU  - Bashir M
AD  - Department of Immunology, Armed Forces Institute of Pathology, 46000 Rawalpindi, 
      Pakistan.
FAU - Tipu, Hamid Nawaz
AU  - Tipu HN
AD  - Department of Immunology, Armed Forces Institute of Pathology, 46000 Rawalpindi, 
      Pakistan.
FAU - Hussain, Mohammad
AU  - Hussain M
AD  - Department of Immunology, Armed Forces Institute of Pathology, 46000 Rawalpindi, 
      Pakistan.
LA  - eng
PT  - Journal Article
DEP - 20180201
PL  - England
TA  - Biomed Rep
JT  - Biomedical reports
JID - 101613227
PMC - PMC5838303
OTO - NOTNLM
OT  - ankylosing spondylitis
OT  - high-resolution polymerase chain reaction
OT  - human leukocyte antigen B27
OT  - sequence specific primers
OT  - spondyloarthropathies
EDAT- 2018/03/16 06:00
MHDA- 2018/03/16 06:01
PMCR- 2018/02/01
CRDT- 2018/03/16 06:00
PHST- 2017/09/20 00:00 [received]
PHST- 2017/12/18 00:00 [accepted]
PHST- 2018/03/16 06:00 [entrez]
PHST- 2018/03/16 06:00 [pubmed]
PHST- 2018/03/16 06:01 [medline]
PHST- 2018/02/01 00:00 [pmc-release]
AID - BR-0-0-1055 [pii]
AID - 10.3892/br.2018.1055 [doi]
PST - ppublish
SO  - Biomed Rep. 2018 Apr;8(4):385-390. doi: 10.3892/br.2018.1055. Epub 2018 Feb 1.