Type 1 Diabetes High-Risk HLA Class II Determination by Polymerase Chain Reaction Sequence-Specific Primers

Objectives: This study aimed to investigate the association between HLA alleles and visceral leishmaniasis (VL) in a sample of Iraqi patients. Methods: A total of 30 patients were studied, in addition to 20 age, gender and ethnicity matched controls. All subjects were genotyped by polymerase chain reaction-sequence specific primers (PCR-SSP) method. Results: Â For HLA-class I region (A and B loci), only HLA-A*19 allele showed a significant (P = 0.031) decreased frequency in VL patients as compared with controls (13.3 vs. 45.0%), and such deviation was associated with OR and PF values of 0.19 and 0.37, respectively. At HLA-class II region, HLA-DRB1*03 and HLA-DQB1*02 alleles were significantly (P = 0.020 and 0.013, respectively) increased in VL patients (56.6 vs. 20% and 46.6 vs. 10%, respectively) as compared with controls. The OR of such two positive associations was 5.23 and 7.88, respectively, and the EF value was 0.46 and 0.41, respectively. In contrast, HLA-DRB1*02 (13.3 vs. 45.0%) and HLA-DQB1*03 (33.3 vs. 70.0%) were significantly (P = 0.031 and 0.023, respectively) decreased in patients. However, none of these differences remained significant after correcting the P value for the number of alleles tested at each locus. Conclusion: these preliminary data suggest that HLA alleles may have some role in aetiopathogenesis of VL, and this role can be in favour of susceptibility and/or protection.

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Nested Polymerase Chain Reaction With Sequence-Specific Primers Typing for HLA-A, -B, and -C Alleles: Detection of Microchimerism in DR-Matched Individuals

Blood ◽

10.1182/blood.v94.4.1471 ◽

1999 ◽

Vol 94 (4) ◽

pp. 1471-1477 ◽

Cited By ~ 16

Author(s):

Anthony S. Carter ◽

Lucia Cerundolo ◽

Mike Bunce ◽

Dicken D.H. Koo ◽

Kenneth I. Welsh ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Nested Pcr ◽

Solid Organ Transplantation ◽

Class Ii ◽

Hla Class Ii ◽

Solid Organ ◽

Chain Reaction ◽

Specific Primers ◽

Polymerase Chain ◽

Hla Class Ii Alleles

Abstract It is widely accepted that donor leukocytes survive within the recipient periphery after blood transfusion or solid organ transplantation. The significance of this microchimerism remains unclear, partially because of the insecurity of assays used to detect the donor-derived material. The techniques used to detect donor-derived DNA within recipient peripheral blood rely largely on major histocompatibility complex class II polymorphism. We and others have shown that the sensitivity of polymerase chain reaction with sequence-specific primers (PCR-SSP) typing for HLA class II alleles can be increased 100-fold by the addition of a primary amplification step (nested PCR-SSP). We have now extended this technique to encompass typing for HLA class I alleles, thereby adding flexibility to microchimerism testing by enabling testing of recipients HLA-DR matched with their donors. However, the high level of sensitivity achieved with the technique (1:100,000) leads to a concomitant decrease in the specificity that results in the amplification of unexpected products, a phenomenon we encountered in the development of our nested PCR-SSP typing system for HLA class II alleles. We describe here how it is possible to compensate for these anomalies by including multiple testing of a pretransfusion sample that acts as a specificity control, establishing a rigorous baseline for subsequent analysis.

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Nested Polymerase Chain Reaction With Sequence-Specific Primers Typing for HLA-A, -B, and -C Alleles: Detection of Microchimerism in DR-Matched Individuals

Blood ◽

10.1182/blood.v94.4.1471.416k10_1471_1477 ◽

1999 ◽

Vol 94 (4) ◽

pp. 1471-1477

Author(s):

Anthony S. Carter ◽

Lucia Cerundolo ◽

Mike Bunce ◽

Dicken D.H. Koo ◽

Kenneth I. Welsh ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Nested Pcr ◽

Solid Organ Transplantation ◽

Class Ii ◽

Hla Class Ii ◽

Solid Organ ◽

Chain Reaction ◽

Specific Primers ◽

Polymerase Chain ◽

Hla Class Ii Alleles

It is widely accepted that donor leukocytes survive within the recipient periphery after blood transfusion or solid organ transplantation. The significance of this microchimerism remains unclear, partially because of the insecurity of assays used to detect the donor-derived material. The techniques used to detect donor-derived DNA within recipient peripheral blood rely largely on major histocompatibility complex class II polymorphism. We and others have shown that the sensitivity of polymerase chain reaction with sequence-specific primers (PCR-SSP) typing for HLA class II alleles can be increased 100-fold by the addition of a primary amplification step (nested PCR-SSP). We have now extended this technique to encompass typing for HLA class I alleles, thereby adding flexibility to microchimerism testing by enabling testing of recipients HLA-DR matched with their donors. However, the high level of sensitivity achieved with the technique (1:100,000) leads to a concomitant decrease in the specificity that results in the amplification of unexpected products, a phenomenon we encountered in the development of our nested PCR-SSP typing system for HLA class II alleles. We describe here how it is possible to compensate for these anomalies by including multiple testing of a pretransfusion sample that acts as a specificity control, establishing a rigorous baseline for subsequent analysis.

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Insulin Administration May Trigger Type 1 Diabetes in Japanese Type 2 Diabetes Patients With Type 1 Diabetes High-Risk HLA Class II and the Insulin Gene VNTR Genotype

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2014-1759 ◽

2014 ◽

Vol 99 (9) ◽

pp. E1793-E1797 ◽

Cited By ~ 3

Author(s):

Wataru Nishida ◽

Masao Nagata ◽

Akihisa Imagawa ◽

Toshiaki Hanafusa ◽

Jun Ohashi ◽

...

Keyword(s):

Type 2 Diabetes ◽

Type 1 Diabetes ◽

High Risk ◽

Class Ii ◽

Hla Class Ii ◽

Insulin Gene ◽

Insulin Administration ◽

Diabetes Patients

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DRB Genotyping in Cynomolgus Monkeys from China Using Polymerase Chain Reaction -Sequence-Specific Primers

Human Immunology ◽

10.1016/j.humimm.2006.10.010 ◽

2007 ◽

Vol 68 (2) ◽

pp. 135-144 ◽

Cited By ~ 7

Author(s):

Huafeng Wei ◽

Hao Wang ◽

Sheng Hou ◽

Shuiqing Hu ◽

Kexing Fan ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Reaction Sequence ◽

Chain Reaction ◽

Specific Primers ◽

Cynomolgus Monkeys ◽

Polymerase Chain

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HLA class II alleles and haplotypes in Lithuanian children with type 1 diabetes and healthy children (HLA and type 1 diabetes)

Medicina ◽

10.3390/medicina46080072 ◽

2010 ◽

Vol 46 (8) ◽

pp. 505 ◽

Cited By ~ 5

Author(s):

Erika Skrodenienė ◽

Dalia Marčiulionytė ◽

Žilvinas Padaiga ◽

Edita Jašinskienė ◽

Vaiva Sadauskaitė-Kuehne ◽

...

Keyword(s):

Diabetes Mellitus ◽

Type 1 Diabetes ◽

Type 1 Diabetes Mellitus ◽

Clinical Manifestations ◽

Class Ii ◽

Hla Class Ii ◽

Healthy Children ◽

Polymerase Chain ◽

Control Study

Objective. Type 1 diabetes mellitus is a slowly progressive autoimmune disease. The genetic background of type 1 diabetes mellitus is polygenic with the major disease locus located in the human leukocytes antigen (HLA) region. High risk and protective alleles, haplotypes, and genotypes have been determined in Lithuanian children with type 1 diabetes mellitus and healthy children. Material and methods. In this case-control study, 124 children with diabetes (55 males and 69 females; mean age, 9.2±3.9 years) were tested for HLA class II and compared with 78 healthy controls (43 males and 35 females; mean age, 10.8±3.4 years; range, 0–15 years). HLA DRB1, DQA1, and DQB1 alleles were genotyped using a polymerase chain reaction. Results. T1D risk-associated haplotypes (DR4)-DQA1*0301-DQB1*0302, (DR3)- DQA1*0501-DQB1*0201, and (DR1)-DQA1*0101-04-DQB1*0501 were more prevalent among children with diabetes than controls (50.0%, 41.1%, and 37.9% vs. 10.3%, 5.1%, and 24.4%, P<0.001). The haplotypes (DR4)-DQA1*0301-DQB1*0302 and (DR3)-DQA1*0501-DQB1*0201 increased T1D risk by 8.75 and 12.93 times, respectively (P<0.001). Protective haplotypes (DR2)- DQA1*0102-B1*0602, (DR11/12/13)-DQA1*05-DQB1*0301, and (DR13)-DQA1*0103- DQB1*0603 were significantly more prevalent among controls than children with diabetes (25.6%, 33.3%, 19.2% vs. 0%, 3.2%, 0%; P<0.001). These frequencies are quite similar to those from neighbor countries with varying incidence of type 1 diabetes mellitus. Conclusions. HLA class II haplotypes associated with type 1 diabetes mellitus positively or negatively were the same in Lithuanian children as in other European Caucasian populations. Differences in incidence and clinical manifestations of type 1 diabetes might be due to different environmental factors and/or lifestyle.

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