PMID- 10932047
OWN - NLM
STAT- MEDLINE
DCOM- 20000918
LR  - 20061115
IS  - 0003-3898 (Print)
IS  - 0003-3898 (Linking)
VI  - 58
IP  - 4
DP  - 2000 Jul-Aug
TI  - [Standardization and automation of HLA B27 typing by flow cytometry: validation 
      and comparison with microlymphocytotoxicity].
PG  - 461-6
AB  - One of the strongest known association between human leukocyte antigen (HLA) 
      phenotype and disease is that of ankylosing spondylitis and HLA-B27. Thus, the 
      determination of HLA-B27 status is an useful tool in the diagnosis of ankylosing 
      spondylitis. To date, the 2 reference methods for HLA typing 
      (microlymphocytotoxicity and molecular biology techniques), are costly in terms 
      of both technician time and materials, and require a great deal of experience. In 
      total, these techniques are not well-suited for routine application in clinical 
      immunology laboratories. Use of flow cytometry has recently been applied for 
      HLA-B27 typing. Nevertheless, it requires an extensive validation protocol. We 
      developed a flow cytometry technique as standardized as possible (whole blood, 
      automated lysing system, automated photomultiplier voltage calibration, 
      definition of thresholds stable with time) and validated our results by 
      comparison with microlymphocytotoxicity. In total, 326 samples were analyzed. We 
      found 99% of concordant results between the 2 techniques, and neither false 
      positive results nor false negative results with flow cytometry could be 
      observed. These results illustrate the reliability of the protocol. It should be 
      remembered that reference technique remains necessary to confirm the few results 
      (< 1%) found in "grey zone" by flow cytometry. Standardization of flow cytometry 
      techniques, as described in this work for HLA B27, seems to be a reasonable goal 
      for the next decade in clinical immunology laboratories.
FAU - Monneret, G
AU  - Monneret G
AD  - Laboratoire d'immunologie, Centre hospitalier Lyon-Sud.
FAU - Seffert, O
AU  - Seffert O
FAU - Debard, A L
AU  - Debard AL
FAU - Gutowski, M C
AU  - Gutowski MC
FAU - Couprie, N
AU  - Couprie N
FAU - Larbre, J P
AU  - Larbre JP
FAU - Tebib, J
AU  - Tebib J
FAU - Bienvenu, J
AU  - Bienvenu J
LA  - fre
PT  - Comparative Study
PT  - English Abstract
PT  - Journal Article
TT  - Standardisation et automatisation du typage HLA B27 par une technique 
      cytofluorimetrique: validation par comparaison a la microlymphocytoxicite.
PL  - France
TA  - Ann Biol Clin (Paris)
JT  - Annales de biologie clinique
JID - 2984690R
RN  - 0 (HLA-B27 Antigen)
SB  - IM
MH  - Automation/methods/standards
MH  - Cytotoxicity Tests, Immunologic/*methods
MH  - Female
MH  - Flow Cytometry/*methods/standards
MH  - HLA-B27 Antigen/*blood/genetics
MH  - Histocompatibility Testing/*methods/standards
MH  - Humans
MH  - Male
MH  - Middle Aged
MH  - Phenotype
MH  - Quality Control
MH  - Spondylitis, Ankylosing/diagnosis/immunology
EDAT- 2000/08/10 11:00
MHDA- 2000/09/23 11:01
CRDT- 2000/08/10 11:00
PHST- 2000/08/10 11:00 [pubmed]
PHST- 2000/09/23 11:01 [medline]
PHST- 2000/08/10 11:00 [entrez]
PST - ppublish
SO  - Ann Biol Clin (Paris). 2000 Jul-Aug;58(4):461-6.