PMID- 36377987
OWN - NLM
STAT- MEDLINE
DCOM- 20221118
LR  - 20221118
IS  - 1433-6510 (Print)
IS  - 1433-6510 (Linking)
VI  - 68
IP  - 11
DP  - 2022 Nov 1
TI  - Comparative Analysis of a Developed Probe-Based Real-Time PCR and PCR-SSP for 
      HLA-B*27 Detection among Thai Blood Donors.
LID - 10.7754/Clin.Lab.2022.220231 [doi]
AB  - BACKGROUND: The human leukocyte antigen (HLA) class I gene, the B locus, allele 
      27, HLA-B*27 is one of the most fascinating risk factors that is strongly 
      associated with developing spondyloarthropathies (SpA). HLA-B27 testing has been 
      routinely available in the diagnosis of those diseases. This study aimed to 
      develop a fluorogenic real-time PCR and to compare it with PCR-SSP to detect the 
      HLA-B*27 allele among Thai blood donors. METHODS: A total of 391 DNA samples were 
      obtained from Thai blood donors at Thammasat University Hospital and tested for 
      HLA-B*27 allele detection. A new real-time PCR was developed and validated to 
      identify this allele and subsequently compared with those results tested with 
      PCR-SSP. The sensitivity of detection was performed using known HLA-B*27-positive 
      and -negative samples with concentrations ranging from 0.001 to 100 ng/microL. 
      Additionally, HLA-B27 subtyping was performed by DNA sequencing containing second 
      and third exons of this gene among all the HLA-B*27-positive donors. RESULTS: The 
      validity of real-time PCR using known DNA controls and the results obtained by 
      PCR-SSP techniques were in 100% concordance. The method was sensitive even at low 
      DNA concentrations (1 ng/microL). Of 391 donors, 24 (6.14%; 95% CI, 3.97 - 9.00) were 
      found to have the HLA-B*27 allele, while the remaining 367 (93.86%; 95% CI, 91.00 
      - 96.03) did not have this allele. Donors presented HLA-B*27-positive, 
      HLA-B*27:06, the most common allele, followed by HLA-B*27:04, -B*27:05, and 
      -B*27:07. CONCLUSIONS: HLA-B*27 using fluorogenic real-time qualitative PCR was 
      found to be superior compared with that of PCR-SSP. The method is rapid, 
      accurate, reliable, and sensitive for detection. In addition, this method 
      provides convenience in the early treatment of SpA patients and relieves their 
      suffering.
FAU - Intharanut, Kamphon
AU  - Intharanut K
FAU - Ruanthong, Fonthip
AU  - Ruanthong F
FAU - Chidtrakoon, Sarisa
AU  - Chidtrakoon S
FAU - Nathalang, Oytip
AU  - Nathalang O
LA  - eng
PT  - Journal Article
PL  - Germany
TA  - Clin Lab
JT  - Clinical laboratory
JID - 9705611
RN  - 0 (HLA-B27 Antigen)
RN  - 9007-49-2 (DNA)
SB  - IM
MH  - Humans
MH  - *HLA-B27 Antigen/genetics/analysis
MH  - Real-Time Polymerase Chain Reaction
MH  - *Blood Donors
MH  - Thailand
MH  - Alleles
MH  - DNA/genetics
EDAT- 2022/11/16 06:00
MHDA- 2022/11/19 06:00
CRDT- 2022/11/15 09:25
PHST- 2022/11/15 09:25 [entrez]
PHST- 2022/11/16 06:00 [pubmed]
PHST- 2022/11/19 06:00 [medline]
AID - 10.7754/Clin.Lab.2022.220231 [doi]
PST - ppublish
SO  - Clin Lab. 2022 Nov 1;68(11). doi: 10.7754/Clin.Lab.2022.220231.