Virtual DNA analysis as a platform for interlaboratory data exchange of HLA DNA typing results.

In 1998, the German DNA Exchange offered the possibility to report typing data as virtual DNA. Participating labs have been equipped with software based on the principle of Virtual DNA Analysis (VDA). This approach allows the combination of sequence-specific oligonucleotide (SSO), sequence-specific primer (SSP) and sequence-based typing (SBT) results. The use of all types of test kits has been allowed without any limitations, as long as basic sequence information on SSOs or SSPs was available, at least the sequence and the position of the detected motif on the sample DNA. Typing raw data of the actual SSO-SSP and, if performed, SBT information was collected. Participating labs received 20 DNA samples to type. Fourteen labs returned data on 1,250 single-locus testings. Reported data consisted of 317 SBT data, 452 SSO kits and 1,795 SSP kits with 43,312 single SSO/ SSP reactivities. One hundred and twenty-six different typing kits (unique laboratory-specific kits, commercial kits from 7 companies) have been used. In 30 (2.4%) single-locus testings, at least one single SSO/SSP reactivity has been false-positive or -negative, thus not leading to a valid result on primary evaluation. Eight of these 30 cases were due to the presence of a new DRB1*14 allele in sample no. 2. Thirty-five tests (2.8%) showed wrong allele assignments. This first attempt to collect raw typing data instead of typing interpretation on a larger scale shows the advantages of Virtual DNA Analysis like interlaboratory data exchange without loss of information, transparency of typing interpretation and reinterpretation of typing data with an updated allele database. The VDA format is a useful tool for workshops and bone marrow donor registries.

[1]  Norman Arnheim,et al.  New HLA–DPB1 alleles generated by interallelic gene conversion detected by analysis of sperm , 1995, Nature Genetics.

[2]  D. Monos,et al.  Large-scale DRB and DQB1 oligonucleotide typing for the NMDP registry: progress report from year 2. , 1996, Tissue antigens.

[3]  W. Helmberg,et al.  Virtual DNA analysis--a new tool for combination and standardised evaluation of SSO, SSP and sequencing-based typing results. , 1998, Tissue antigens.

[4]  C. Hurley,et al.  Novel HLA-B alleles, B*8201, B*3515 and B*5106, add to the complexity of serologic identification of HLA types. , 1996, Tissue antigens.

[5]  S G Marsh,et al.  HLA class II region sequences, 1998. , 2008, Tissue antigens.

[6]  Shirley A. Miller,et al.  A simple salting out procedure for extracting DNA from human nucleated cells. , 1988, Nucleic acids research.

[7]  C. Hurley Acquisition and use of DNA-based HLA typing data in bone marrow registries. , 1997, Tissue antigens.

[8]  P Parham,et al.  HLA class I region sequences, 1998. , 2008, Tissue antigens.

[9]  J. Hansen,et al.  A comprehensive approach for typing the alleles of the HLA-B locus by automated sequencing. , 1995, Tissue antigens.

[10]  C. Nevinny-Stickel,et al.  HLA CLASS II TYPING IN A MICROTITRE PLATE FORMATE USING DIGOXIGENIN‐LABELLED AMPLIFIED DNA AND BIOTIN‐LABELLED OLIGONUCLEOTIDE PROBES , 1993, European journal of immunogenetics : official journal of the British Society for Histocompatibility and Immunogenetics.