HLA-SuBiTo

HLA-SuBiTo - the sequencing system

Simple and consistent setup for all HLA-SuBiTo kits
  • To separate as many heterozygous allele combinations in the first step, 2 parallel amplifications are performed
  • gel electrophoresis
  • Purification of PCR products with ExoSAP-IT TM
  • 6 sequencing reactions per HLA class I locus sequencing
  • 2-3 sequencing reactions per HLA class II locus sequencing
  • The combination of forward and reverse reactions occurs depending on the positive PCR amplification
  • Loading the capillary sequencer
  • Analysis with the HLA-HiType software
Saves 3 hours of assay time with kit enhancements
  • New G1 / G2 mix chemistry allows shorter first PCR run time of only 1 h 45 min
  • Shorter cycle sequencing PCR protocol lasts only 1 h 10 min
  • Alternative purification protocol saves 30 minutes
Kit components
  • All components for the amplification incl. IT-Taq polymerase
  • Exo-SAP-IT TM for the purification of PCR products
  • Genort- and Expon-specific sequencing mixes incl. BigDye ® chemistry
  • IT Prec buffer for precipitation of sequencing products
  • Two second level sequencing mixes for the resolution of frequent ambiguities.
item number Surname Test / plate
RUO Emergency for use in diagnostic procedures - USA and Canada
002 090 025 HLA-SuBiTo A 25
002 090 025/1 HLA-SuBiTo A without BDT's 25
002 091 025 HLA-SuBiTo B 25
002 091 025/1 HLA-SuBiTo B without BDT's 25
002 092 025 HLA-SuBiTo C 25
002 092 025/1 HLA-SuBiTo C without BDT's 25
002 093 025 HLA-SuBiTo DRB1 25
002 093 025/1 HLA-SuBiTo DRB1 without BDT's 25
002 094 025 HLA-SuBiTo DQB1 25
002 094 025/1 HLA-SuBiTo DQB1 without BDT's 25
002 095 025 HLA-SuBiTo DPB1 25
002 095 025/1 HLA-SuBiTo DPB1 without BDT's 25
002 096 025 HLA-SuBiTo ABCDRDQ 25
002 096 025/1 HLA-SuBiTo ABCDRDQ without BDT's 25


Second level sequencing

Due to the HLA-SuBiTo strategy, which separates HLA allele groups with two initial PCR amplifications, the common cis / trans ambiguities are generally greatly reduced compared to single-amplification strategies. In the event that HLA allele groups are not separated, possible ambiguities can be resolved with our ready-to-use second-level sequencing mixes and the existing standard kit PCR product.

A total of 16 different second-level sequencing mixes are currently available for HLA class I and six each for DRB1 * and DQB1.

The HLA-HiType software shows in ambiguities, with which second level sequencing mix these can be resolved.

Evaluation with the HLA-HiType software
  • New nomenclature and automatic conversion into P and G groups as well as new NMDP codes integrated
  • Fully automatic data management, tedious selection of * .ab1 files is not necessary
  • Automatic recognition and dynamic representation of hemi- and heterozygous sequences
  • Various manual and automatic editing options
  • Different alignment options, combined control of electropherogram and alignment
  • High level of safety through a defined 4-eyes principle (technical and medical validation)
  • Complete history and traceability