Development of a new HLA-DRB real-time PCR typing method

Casamitjana, Natàlia; Faner, Rosa; Balaguer, Albert; Colobran, Roger; Pujol-Borrell, Ricardo; Juan, Manel; Palou, Eduard

doi:10.1016/j.humimm.2004.08.178

Cited by 14 publications

(8 citation statements)

References 12 publications

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“…Sequence‐specific primers (SSP), sequence‐specific oligonucleotide probes (SSOP), sequencing‐based typing (SBT), polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP), polymerase chain reaction–single‐strand conformational polymorphism (PCR‐SSCP) and reference strand‐mediated conformation analysis (RSCA) were molecular methods traditionally and currently used for MHC typing (1–12). Improvement of high‐throughput methods based on sequencing, pyrosequencing, real‐time PCR, TaqMan assay and microarrays have been developed in recent years (13–17).…”

Section: Ela‐dra Allele Frequencies In Domestic Horse Breeds the Allmentioning

confidence: 99%

Development of an ELA‐DRA gene typing method based on pyrosequencing technology

Díaz

Echeverria

et al. 2008

Tissue Antigens

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The polymorphism of equine lymphocyte antigen (ELA) class II DRA gene had been detected by polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP) and reference strand-mediated conformation analysis. These methodologies allowed to identify 11 ELA-DRA exon 2 sequences, three of which are widely distributed among domestic horse breeds. Herein, we describe the development of a pyrosequencing-based method applicable to ELA-DRA typing, by screening samples from eight different horse breeds previously typed by PCR-SSCP. This sequence-based method would be useful in high-throughput genotyping of major histocompatibility complex genes in horses and other animal species, making this system interesting as a rapid screening method for animal genotyping of immune-related genes.

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Section: Ela‐dra Allele Frequencies In Domestic Horse Breeds the Allmentioning

confidence: 99%

Development of an ELA‐DRA gene typing method based on pyrosequencing technology

Díaz

Echeverria

et al. 2008

Tissue Antigens

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“…[Slateva et al, 2001;Sylvain et al, 2004;Casamitjana et al, 2005]) and one report of a high resolution TaqMan-based typing assay [Tremmel et al, 1999]. In contrast to our study, Tremmel et al (1999) relied largely on sequence-specific primers to discriminate alleles of DRB1*15 and *16 and incorporated a separate internal control amplification into each reaction.…”

Section: Discussionmentioning

confidence: 90%

A real‐time PCR approach for rapid high resolution subtyping of HLA‐DRB1*04

Gersuk

Nepom

2006

Journal of Immunological Methods

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We present a real-time PCR approach for the identification and subtyping of HLA-DR4 alleles. The technique, which uses sequence-specific primers and probes in conjunction with real-time PCR for the detection and differentiation of target alleles, is rapid, involves minimal hands-on time, and is inexpensive compared to existing methods. Further, there is no post-PCR handling, so the risk of contamination is avoided. We have validated the assay using 44 blinded and 56 unblinded samples, which were identified with 100% accuracy, sensitivity, and specificity. We demonstrate the applicability of this assay as an alternative approach to traditional HLA typing methods.

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“…In the past, many HLA typing methods were developed and applied for transplantation purposes. These methods are divided into serological 40 and DNA-based typing commonly [16][17][18][19] . However, due to the cross-reactive nature of the alloantibody, serological typing methods such as complement-dependent give misleading results 41 .…”

Section: Allele Assignmentmentioning

confidence: 99%

“…Due to the unique characteristics of each technique, various DNAbased typing methods have been used independently or in combination with each other for transplantation purposes. However, previous DNA-based methods are somewhat limited in that typing reactions are required for a large number of samples [16][17][18][19] . To resolve this problem, microarray technology has been applied for HLA genotyping of HLA-A 22 , HLA-B 20 , and HLA-DRB1 21,22 .…”

Section: Allele Assignmentmentioning

confidence: 99%

“…This method is simple compared to HLA genotyping, and more samples can be applied compared to the PCR-SSPs method. Sequence-based typing (SBT) can be used to perform high resolution genotyping and detect new alleles 18 , and real-time PCR is recognized as a quick HLA genotyping method 19 . However, these methods are somewhat limited due to their requirements for a large number of samples and complex experimental procedures.…”

Section: Introductionmentioning

confidence: 99%

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Development of simple and rapid HLA-C genotyping method using an oligonucleotide microarray

et al. 2011

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Human Leukocyte Antigen (HLA), called human Major Histocompatibility Complex (MHC), performs the crucial function of antigen presentation in human immunity, and correctly matched HLA antigens improves survival and reduces immunological complications. Therefore, a precise HLA typing method is necessary for successful transplantation between donor and recipient. Until now, many methods have been developed for HLA typing, including serological and DNA-based methods. However, these previous methods have been somewhat limited due to their requirements for a large number of samples and complex experimental procedures. To resolve this problem, in this study, we developed rapid genotyping methods for HLA-C using an oligonucleotide microarray. To evaluate the performance of the HLA-C genotyping microarray, we analyzed 50 unrelated Korean individuals. The genotyping results obtained by the oligonucleotide microarray were identical with those of PCR-SBT and PCR-SSPs, at a low-resolution, 2-digit level. Therefore, this microarray method for genotyping HLA constitutes a rapid, simultaneous, and high-throughput method that can be used as an alternative to current methods.

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Development of a new HLA-DRB real-time PCR typing method

Cited by 14 publications

References 12 publications

Development of an ELA‐DRA gene typing method based on pyrosequencing technology

Development of an ELA‐DRA gene typing method based on pyrosequencing technology

A real‐time PCR approach for rapid high resolution subtyping of HLA‐DRB1*04

Development of simple and rapid HLA-C genotyping method using an oligonucleotide microarray

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