Skip to main content
NCBI home page
Journal of Clinical Pathology logoLink to Journal of Clinical Pathology
. 1995 Jan;48(1):33–36. doi: 10.1136/jcp.48.1.33

HLA typing for DR3 and DR4 using artificial restriction fragment length polymorphism PCR from archival DNA.

V A Horton 1, M Bunce 1, D R Davies 1, R C Turner 1, Y M Lo 1
PMCID: PMC502258  PMID: 7706515

Abstract

AIM--To develop polymerase chain reaction based artificial restriction fragment length polymorphism (artificial RFLP PCR) assays for DR3 and DR4 alleles of the multiallelic DRB1 locus and to apply them to paraffin wax embedded archival material. METHODS--Sixty five samples from DRB1 typed cell lines were analysed using the artificial RFLP PCR method to determine the specificity and sensitivity of the system. RESULTS--The artificial RFLP PCR method for typing the DRB1 locus showed 100% accuracy in the 65 samples previously typed using allele specific PCR and serology. The samples included 18 combinations of alleles that included DR3, 18 that included DR4, four that were DR3/DR4 heterozygotes, and 10 samples that were neither DR3 nor DR4. Typing of 10 paraffin wax embedded samples using artificial RFLP PCR was in complete agreement with previous typing at the DRB1 locus. CONCLUSION--The application of artificial RFLP PCR for the analysis of multiallelic loci, such as those of the HLA system, in archival DNA samples has been achieved. Artificial RFLP PCR is a robust, easily implemented, non-isotopic system and may be useful for large retrospective studies.

Full text

PDF
33

Images in this article

35Image
on p.35

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. An S. F., Franklin D., Fleming K. A. Generation of digoxigenin-labelled double-stranded and single-stranded probes using the polymerase chain reaction. Mol Cell Probes. 1992 Jun;6(3):193–200. doi: 10.1016/0890-8508(92)90016-q. [DOI] [PubMed] [Google Scholar]
  2. Bein G., Gläser R., Kirchner H. Rapid HLA-DRB1 genotyping by nested PCR amplification. Tissue Antigens. 1992 Feb;39(2):68–73. doi: 10.1111/j.1399-0039.1992.tb01909.x. [DOI] [PubMed] [Google Scholar]
  3. Bidwell J. L., Bidwell E. A., Savage D. A., Middleton D., Klouda P. T., Bradley B. A. A DNA-RFLP typing system that positively identifies serologically well-defined and ill-defined HLA-DR and DQ alleles, including DRw10. Transplantation. 1988 Mar;45(3):640–646. doi: 10.1097/00007890-198803000-00027. [DOI] [PubMed] [Google Scholar]
  4. Bunce M., Taylor C. J., Welsh K. I. Rapid HLA-DQB typing by eight polymerase chain reaction amplifications with sequence-specific primers (PCR-SSP). Hum Immunol. 1993 Aug;37(4):201–206. doi: 10.1016/0198-8859(93)90502-r. [DOI] [PubMed] [Google Scholar]
  5. Chapman R. W. Role of immune factors in the pathogenesis of primary sclerosing cholangitis. Semin Liver Dis. 1991 Feb;11(1):1–4. doi: 10.1055/s-2008-1040415. [DOI] [PubMed] [Google Scholar]
  6. Kimura A., Dong R. P., Harada H., Sasazuki T. DNA typing of HLA class II genes in B-lymphoblastoid cell lines homozygous for HLA. Tissue Antigens. 1992 Jul;40(1):5–12. doi: 10.1111/j.1399-0039.1992.tb01951.x. [DOI] [PubMed] [Google Scholar]
  7. Lo E. S., Lo Y. M., Tse C. H., Fleming K. A. Detection of hepatitis B pre-core mutant by allele specific polymerase chain reaction. J Clin Pathol. 1992 Aug;45(8):689–692. doi: 10.1136/jcp.45.8.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lo Y. M., Lo E. S., Mehal W. Z., Sampietro M., Fiorelli G., Ronchi G., Tse C. H., Fleming K. A. Geographical variation in prevalence of hepatitis B virus DNA in HBsAg negative patients. J Clin Pathol. 1993 Apr;46(4):304–308. doi: 10.1136/jcp.46.4.304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lo Y. M., Mehal W. Z., Fleming K. A. In vitro amplification of hepatitis B virus sequences from liver tumour DNA and from paraffin wax embedded tissues using the polymerase chain reaction. J Clin Pathol. 1989 Aug;42(8):840–846. doi: 10.1136/jcp.42.8.840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Lo Y. M., Mehal W. Z., Wordsworth B. P., Chapman R. W., Fleming K. A., Bell J. I., Wainscoat J. S. HLA typing by double ARMS. Lancet. 1991 Jul 6;338(8758):65–66. doi: 10.1016/0140-6736(91)90067-y. [DOI] [PubMed] [Google Scholar]
  11. Marsh S. G., Bodmer J. G. HLA class II nucleotide sequences, 1992. Tissue Antigens. 1992 Nov;40(5):229–243. doi: 10.1111/j.1399-0039.1992.tb02050.x. [DOI] [PubMed] [Google Scholar]
  12. Newton C. R., Graham A., Heptinstall L. E., Powell S. J., Summers C., Kalsheker N., Smith J. C., Markham A. F. Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS). Nucleic Acids Res. 1989 Apr 11;17(7):2503–2516. doi: 10.1093/nar/17.7.2503. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Olerup O., Zetterquist H. HLA-DR typing by PCR amplification with sequence-specific primers (PCR-SSP) in 2 hours: an alternative to serological DR typing in clinical practice including donor-recipient matching in cadaveric transplantation. Tissue Antigens. 1992 May;39(5):225–235. doi: 10.1111/j.1399-0039.1992.tb01940.x. [DOI] [PubMed] [Google Scholar]
  14. Patel P., Lo Y. M., Bell J. I., Wainscoat J. S. Detection of susceptibility alleles to insulin-dependent diabetes mellitus at the DQB1 locus by artificial PCR-RFLP. Immunogenetics. 1992;36(4):264–265. doi: 10.1007/BF00215057. [DOI] [PubMed] [Google Scholar]
  15. Seki T., Kiyosawa K., Inoko H., Ota M. Association of autoimmune hepatitis with HLA-Bw54 and DR4 in Japanese patients. Hepatology. 1990 Dec;12(6):1300–1304. doi: 10.1002/hep.1840120609. [DOI] [PubMed] [Google Scholar]
  16. Thomson G., Robinson W. P., Kuhner M. K., Joe S., MacDonald M. J., Gottschall J. L., Barbosa J., Rich S. S., Bertrams J., Baur M. P. Genetic heterogeneity, modes of inheritance, and risk estimates for a joint study of Caucasians with insulin-dependent diabetes mellitus. Am J Hum Genet. 1988 Dec;43(6):799–816. [PMC free article] [PubMed] [Google Scholar]
  17. Wolf E., Spencer K. M., Cudworth A. G. The genetic susceptibility to type 1 (insulin-dependent) diabetes: analysis of the HLA-DR association. Diabetologia. 1983 Apr;24(4):224–230. doi: 10.1007/BF00282704. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Pathology are provided here courtesy of BMJ Publishing Group

RESOURCES