Software is used to assign positive or negative reactions based on the strength of the fluorescent signal. Hla typing bioinformatics tools wes analysis omicx. Molecular techniques enabled more accurate analyses of the influence. Many highthroughput molecular hla typing approaches are unable to determine the phase of observed dna sequence polymorphisms, leading to ambiguous results. Principles of hla typing the wakflow hla typing kit is based on the reverse sequencespecific oligonucleotide probes sso method coupled with xmap technology designed for use with. For highthroughput histocompatibility applications such as hla typing of transplant candidates, conventional wisdom suggests sso with luminex provides a fair balance of productivity, turnaround time tat and cost as long as the laboratory can wait to receive sufficient samples to batch them together in one run. Luminex 200 flow cytometry system is a good choice for hla typing.
The use of higher resolution methods is often restricted due to cost and time limitations. Software is used to assign positive or negative reactions based on the strength. Complete hla typing report is delivered upon purchase for each donor 4 digit resolution typing e. Jul 21, 2009 however, for highresolution sso typing of the highly polymorphic hla a or hla b genes this strategy had to be extended. Labtype sso is a costeffective reverse sso dna typing system that provides a complete solution for hla a, b, c. However, current molecular typing methods only interrogate the antigen recognition site ars of hla genes, resulting in many cistrans ambiguities that require additional typing methods to resolve. Luminex xmap technology is an effective strategy for high. Labtype sso dna typing tests for use with labscan3d device using luminex technology for molecular typing and is a tool used in preliminary clinical testing. Unique probe design minimizes ambiguities master mix includes primers, dntps and pcr buffer, eliminating extra steps and consumables. Feb 03, 2017 luminex xmap technology makes up the basis of luminexs core business. Allelelevel resolution data at primary hla typing is the ideal for most histocompatibility testing laboratories. Hla typing by sequencespecific oligonucleotides probes pcr. Several molecular techniques have been adapted for the human leukocyte antigen hla typing with their advantages and drawbacks, which greatly improved the feasibility and precision of hla typing as compared to previously used methods. The labtype sso one lambda, inc and genprobe lifecodes hlasso hla typing tests are rapid.
Hla typing for transplant market global forecast to 2025. Software is used to assign positive or negative reactions based on the strength of the. Hla fusion analyses the data and will place suggested hla typing pairs in the left hand area, which is either accepted or the data manually manipulated by the operator. Hla typing for transplant market by technology pcr sso, ssp. Human leukocyte antigen hla typing, utilising the sequencespecific oligonucleotide sso and sequencespecific primer ssp technologies, has been in routine use in many tissue typing laboratories worldwide for more than 20 years since the development of the polymerase chain reaction. Mar 25, 2015 all on a single assay, a single technology, and a single software platform, the trusight hla panel delivers unambiguous, phaseresolved, ultrahighresolution hla typing for 11 loci. Hla genotyping and antibody characterization using the luminex. The hla typing process then requires a postamplification step to. Expedited hla genotyping can be performed in as little as 10 working days. A key element of the dnabased hla typing procedure is in the design of the primers used for pcr amplification. A suspension array platform employs fluorescently coded microspheres as a solid sup port to immobilize oligonucleotide probes.
Luminex sso hla typing uses sso probes for distinct sequence motifs which are bound to a set of microbeads. The hla typing and serological equivalent assignment area. Hla genotyping and antibody characterization using the. Our diagnostic tests are used to determine donor compatibility pretransplant, and posttransplant to monitor hla antibodies that can lead to rejection. Global hla typing for transplant industry 2019 to 2025. Labtype applies luminex technology to the reverse sso dna typing method. At the top of the left hand section are tabsthese include. We describe here a single antigen bead panel for accurate identification of human leukocyte antigen hla antibody specificities by flow cytometry. The combination of pcr technology and hybridization with sequencespecific oligonucleotide probes was first applied to hla class ii typing because of the limitations of dr serology and of the better knowledge of allelic. Hla genotyping pcrrsso hla next generation hla sequencing. Target dna is amplified with biotinylated, locus specific primers. As the leader in hla for over 30 years, we develop and distribute a comprehensive line of hla typing and antibody detection products, along with instrumentation and software that are used to simplify and automate testing procedures and final test evaluations.
As the leader in hla for over 30 years, we develop and distribute a comprehensive line of hla typing and antibody detection products, along with instrumentation and software that are used to simplify and automate testing procedures and final. The system has already been applied to lowresolution typing e. Ssoluminex, a comparison of high throughput hla typing methods. Luminex 200multiplexing instrument from luminex selectscience. Luminexbased methods in highresolution hla typing request. Easy, fast and good hlasso typing results which are compatible with ssp detection. Mar 30, 2020 chicago, march 30, 2020 prnewswire according to the new market research report hla typing for transplant market by technology pcr sso, ssp, real time, sequencing ngs, sanger, product instrument, reagent, software, application chimerism, antibody screening, end user hospital, diagnolab, academia global forecast to 2025, published by marketsandmarkets, the hla typing for. The performance of a new intermediateresolution method using a pcrluminex platform and labtype sso a, b drb1 kits as an hla typing method for the cord blood cb registry of the korean population was investigated.
Evaluation of labtype sso hla typing using the luminex. Our multigene approach provides hla genotypes for all classical hla genes in one single reaction. For the latest test kits, the companies now use more than 200 different probes on the initial set of 100 luminex microbeads for highresolution hla class i typing. Luminex xmap technology makes up the basis of luminex s core business. Hla typing is done using latest luminex xmap technology based on the principle of hybridization of labeled single stranded pcr product to sso probes attached to luminex microspheres which are read by the compact luminex flowcytometer. Background unambiguous hla typing is important in hematopoietic stem cell transplantation hsct, hla disease association studies, and solid organ transplantation. Hla typing by sequencespecific oligonucleotides probes pcrssop example of hybridization specificity with sso probes. Labtype bead mix, locus specific primer set, primer set dmix, denaturation buffer, neutralization buffer, hybridization buffer, sape buffer, wash buffer. The lifecodes hla typing kits utilize sso methodology to identify hla alleles present in a locusspecific pcramplified sample.
A biotinylated pcr product binds and is subsequently stained by sape. Luminex technology for hla typing by pcr sso and identification of hla antibody specificities. While nextgeneration sequencing data are often available for a patient, deducing the hla genotype is difficult because of substantial sequence similarity within the cluster and exceptionally high variability of the loci. The human leukocyte antigen hla gene cluster plays a crucial role in adaptive immunity and is thus relevant in many biomedical applications. Labtype xr and cwd dna typing and the 2 predicate devices use luminex technology for molecular typing and both are tools used in preliminary clinical testing. With labtype sso you get superior resolution and a costeffective solution for hla a, b, c drb1, drb3, 4, 5, and dqa1,dqb1, dpa1,dpb1 hla typing. Apr 09, 2020 the hla typing for transplant market by technology pcr sso, ssp, real time, sequencing ngs, sanger, product instrument, reagent, software, application chimerism, antibody screening, end. This flexible technology is centered on the use of luminex. An appropriate strategy for class i molecular typing for unos laboratories should at least include generic level typing for all hla a and b types, typing for bw4 and bw6 epitopes, and typing for allelic subgroups of b15 and b40 that can distinguish the common alleles defining the serological splits. Please select either a category andor a product line before searching for documents.
Antibody profiling by solid phase assays is a beadbased, multiplexing technology designed for sensitive and accurate identification and monitoring of antihla antibodies in patients serum samples. All on a single assay, a single technology, and a single software platform, the trusight hla panel delivers unambiguous, phaseresolved, ultrahighresolution hla typing for 11 loci. A total of 110 single recombinant hlas, including 34 a locus alleles, 57 b locus alleles, and 19 c locus alleles, were produced by a mammalian expression system. The luminex flow analyser achieves detection by determining the fluorescent. Well suited for low and high throughput labs, labtype sso is based on the reverse sso rsso method. Interpretation of hla typing results for entry into unet optn. Highresolution genotyping of all hla genes requires only 50 ng gdna. Ideal solution for both low volume and high throughput laboratories. Hla typing with sequencespecific oligonucleotide primed pcr. Hla typing using beadbased methods springer nature. The hla alleles are assigned by analysis of the reaction hybridization pattern of the target sample using the wakflow hla typing. We perform highresolution hla genotyping by nextgeneration sequencing. Linkseqs analysis software, suretyper, generated quick and complete data for each.
This flexible technology is centered on the use of luminex xmap microspheres. The sensitivity and specificity of the hla typing procedure is markedly enhanced by pcr amplification of the target sequence corresponding to the region in the hla gene encoding the polymorphism. Luminexbased methods in highresolution hla typing springer. Apr 08, 2020 dublin, april 08, 2020 globe newswire the hla typing for transplant market by technology pcr sso, ssp, real time, sequencing ngs, sanger, product instrument, reagent, software. Application of highthroughput nextgeneration sequencing for. Hla typing for transplant market by technology pcr sso, ssp, real time, sequencing ngs, sanger, product instrument, reagent, software, application chimerism, antibody screening, end user hospital, diagnolab, academia global forecast to 2025. Pdf luminex technology for hla typing by pcrsso and. Three representative labtype sso dna typing assays from one lambda labtype sso drb1 typing test rss02b1, labtype hd class i b locus typing test rssoh1b, and labtype sso dqadqb typing test rss02q were used to compare the luminex flexmap 3d system using the test xponent v.
Hla typing process then requires a postamplification step to discriminate. Article in french cesbrongautier a1, simon p, achard l, cury s, follea g, bignon jd. The entire process is performed in a single well of a 96well pcr plate. Key words hla typing, luminex, pcrsso, labtype, high resolution. Hladpa1b1 sso typing kit 628936 hla null allele sso typing kit 628939 refer to the users manual for match it.