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Occult hepatitis B infection (OBI) is a public health problem in Burkina Faso. OBI represents a risk factor for the development of cirrhosis and hepatocellular carcinoma (HCC). OBI could be due to mutant viruses undetectable by HBsAg assays or a strong suppression of viral replication and gene expression under the pression of the host immune system. To investigate the role of killer cell immunoglobulin-like receptor (KIR) gene polymorphisms in patients with OBI in Burkina Faso compared to healthy and chronic hepatitis B subjects. A total of 286 participants was recruited, including 42 cases of OBI, 110 cases of chronic hepatitis B and 134 HBV negative subjects. SSP-PCR was performed to search for the presence of KIR genes. The HBV viral load was determined by qPCR. The frequencies of the activator gene KIR2DS5 (P=0.045) and the pseudogene KIR2DP1 (P<0.001) in patients with OBI were higher than those in patients with chronic hepatitis B. These genes are associated with susceptibility of occult hepatitis B infection. The frequencies of the inhibitory KIR gene KIR2DL3 (P=0.01) of patients with occult hepatitis B were lower than those in chronic hepatitis B patients. This gene KIR2DL3 is associated with protection against occult hepatitis B infection. Also, the frequencies of the inhibitory KIR genes KIR2DL2 (P<0.001), KIR2DL3 (P<0.001) and activators KIR2DS2 (P<0.001) in chronic hepatitis B patients were higher compared to the frequencies of the KIR genes in healthy subjects. These genes KIR2DL3, KIR2DL5 (A, B), KIR3DL3, KIR3DS1, KIR2DL2 and KIR2DS2 are thought to be genes associated with the susceptibility to OBI. The KIR2DS5 and KIR2DP1 genes could be associated with susceptibility to OBI. As for the KIR gene KIR2DL3 could be associated with protection against occult hepatitis B infection.

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Background and Objectives: Dengue fever (DF), an emerging and re-emerging viral disease, is a major public health problem. The aim of this study was to investigate the influence of KIRs genes polymorphism and KIRs genotypes in susceptibility to dengue virus infection and disease severity in a population from Burkina Faso through a case-control study. Methods: KIRs genes determination was performed using PCR-SSP in 50 patients infected by dengue virus (DENV) and 54 Healthy controls (HC) subjects who had never been infected. Results: Data analysis showed significant association between frequencies of three KIR genes and dengue virus infection (DF): KIR2DL2 (OR: 7.32; IC: 2.87-18.65; P < 0.001); KIR2DL5A (OR: 15.00, IC: 5.68-39.59; P < 0.001) and KIR2DL5B (OR: 11.43; IC: 4.42-29; P < 0.001). While, KIR3DL3 (OR: 0.13, IC: 0.052-0.32; P < 0.001) and KIR2DS5 (OR: 0.12; IC: 0.04-0.30; P < 0.001) were associated with protection against DF. KIR2DL4 (OR: 9.75; IC95%: 1.33-70.97; p: 0.03) and KIRD3DL1 (OR: 12.00; IC95%: 1.60-90.13; p: 0.02) were associated with an increased risk in the development of secondary dengue infection (SDI). Conclusion: The results suggest a contribution of KIR2DL2, KIR2DL5A, and KIR2DL5B genes in the susceptibility of DF development. In contrast, KIR3DL3 and KIR2DS5 were associated with protection against DF development by enhancing both innate and acquired immune responses.

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OBJECTIVE: To investigate Diego blood group alleles in the Chinese Korean population. The Diego blood group system plays an important role in transfusion medicine, but the distribution of the blood group in many Chinese ethnic populations remains unclear. METHODS: Sequence Specific Primer Polymerase Chain Reaction (SSP-PCR) was used for Diego genotyping and sequence-based typing PCR (PCR-SBT) was used to verify single nucleotide polymorphisms in the coding region of SLC4A1 starting from exon 19. Nine hundred and ten samples from the Chinese Korean population were investigated. RESULTS: The frequency of the DI*01 and DI*02 alleles in the Chinese Korean population was 0.0516 and 0.9484, respectively. The most predominant genotype was DI*02/DI*02, with a frequency of 90.22 % (821/910). The frequency of DI*01/DI*02 was 9.23 % (84/910) and that of DI*01 /DI*01 was 0.55 % (5/910). The genotype distributions of the Diego blood group conformed to the Hardy-Weinberg equilibrium (P > 0.05). CONCLUSION: The data obtained from this study will be helpful for the creation of a donor database to provide antigen-negative blood to patients with allo-antibodies. Genotyping can be used as a substitute for the serological technique when antisera are unavailable and is suitable for screening a large number of donors for rare-blood-group databases.

Asunto(s)

Tipificación y Pruebas Cruzadas Sanguíneas/métodos , Alelos , Pueblo Asiatico , Donantes de Sangre , Femenino , Genotipo , Humanos , Masculino , República de Corea

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BACKGROUND: NK cells as a part of innate immune system, are controlled by a set of activating and inhibitory KIR receptors (aKIR, iKIR) which are implicated in tumor microenvironment immunity through a variety of activating and inhibitory immune signals. KIRs are multi gene family receptors that differ in the number and type of genes among individuals. In the current research we determined the KIRs genes and genotypes impact on predisposition to meningioma development in Iranians. METHODS: Sequence-specific primers-polymerase chain reaction (SSP-PCR) was performed for genotyping of 16 KIRs in 159 meningioma cases and 362 age and sex matched healthy controls (CNs) at Shiraz Institute for Cancer Research. RESULTS: Comparison of the KIR genotypes frequencies between cases and controls disclosed a highly significant increase in Bx genotype, CxTx subset and Cen AB and Tel AB in meningioma cases and a decrease in AA genotype, C4Tx subset and Cen AA, Tel AA, Tel BB in healthy controls. Among all 16 KIR genes, the carriers of KIR2DL5 and KIR2DS5 constituted a much greater proportion in meningioma than control group. Comparison of carrier frequencies of KIR2DS4 variants between case and controls revealed a higher frequency of KIR2DS4 full length (KIR2DS4fl) in meningioma cases and a lower frequency of KIR2DS4 deleted variant (KIR2DS4del) in controls. Furthermore, the simultaneous presence of 2DS5, 2DS4fl, CenAB, TelAB and absence of 2DS4del, CenAA, TelAA, TelBB, magnify the risk of developing meningioma substantially (OR ≈ 23). Altogether, 41 distinct KIR genotypes were characterized in 521 subjects. Among them, some individuals were characterized by seven peculiar genotypes that the linkage disequilibrium between KIR2DS2-KIR2DL2 and KIR2DL5-KIR2DS3-KIR2DS5 has not been detected. The carriers of certain genotypes with presence of as KIR2DL5 and absence of KIR2DS3, KIR2DS5 constituted a much higher proportion in meningioma than control group which increase the risk of meningioma up to 72 times. CONCLUSION: This case- control study suggests carriers of Bx genotype, KIR2DL5, KIR2DS5, 2DS4fl, ≥ 4 iKIR, CxTx subset as well as Cen AB and Tel AB are associated with an increased risk of developing meningioma whereas carrying KIR2DS4del, AA, C4TX genotypes and Cen AA, Tel AA, Tel BB reduce the genetic predisposition for meningioma.

Asunto(s)

Neoplasias Meníngeas/genética , Meningioma/genética , Receptores KIR/genética , Estudios de Casos y Controles , Femenino , Frecuencia de los Genes/genética , Genotipo , Humanos , Irán , Células Asesinas Naturales/metabolismo , Masculino , Persona de Mediana Edad , Microambiente Tumoral/genética

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BACKGROUND: A set of activating and inhibitory KIRs (aKIR, iKIR) are involved in NK cell mediated immunity. This study was carried out in order to investigate the KIRs pattern and its association with colorectal carcinoma (CRC) development and clinical outcomes. METHODS: Sequence-specific primers-polymerase chain reaction (SSP-PCR) for typing of 16 KIR genes was utilized in 165 patients with colorectal adenocarcinoma with 165 age and gender matched healthy controls (CNs). RESULTS: Possessing KIR2DS1, 2DS5, 3DS1, 2DS4fl, 2DL5, telomeric half KIR genes, ≥ 4 aKIR and CXT4 genotype were associated with an increased susceptibility to colorectal adenocarcinoma while KIR2DS4del and iKIR >aKIR confer resistance to CRC. On the other hand, clinical associations revealed the defensive role of telomeric KIR3DL1, 3DS1, 2DS1, 2DS4, genotypes with ≥ 4 aKIR and more inhibitory KIRs than activating ones (I > A) against metastasis and CXTX genotype in perineural invasion. CONCLUSION: According to current results it appears that KIRs system play distinctive roles in development and metastasis of colorectal adenocarcinoma.

Asunto(s)

Neoplasias Colorrectales/etiología , Neoplasias Colorrectales/patología , Predisposición Genética a la Enfermedad , Receptores KIR/genética , Anciano , Alelos , Estudios de Casos y Controles , Neoplasias Colorrectales/metabolismo , Femenino , Frecuencia de los Genes , Estudios de Asociación Genética , Genotipo , Humanos , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Masculino , Persona de Mediana Edad , Familia de Multigenes , Metástasis de la Neoplasia , Estadificación de Neoplasias , Receptores KIR/metabolismo

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HLA-A*29 and HLA-B*51 are associated with birdshot uveitis and Behçet's disease, respectively, and are used as a diagnostic criterion in patients with suspected disease, requiring their detection in diagnostic laboratories. While commercial tests for individual HLA alleles are available for other disease-associated HLA variants, no similar allele-specific assays are available for HLA-A*29 and HLA-B*51. Here, we report sequence-specific priming-polymerase chain reaction (SSP-PCR) methods for the detection of HLA-A*29 and HLA-B*51 using a single PCR reaction per allele. The assays were tested in 30 and 32 previously HLA-typed samples, respectively, representing >97% of HLA-A alleles and >93% of HLA-B alleles in a European population. A concordance of 100% was observed with previous typing results, validating these methods for use in a diagnostic or research context.

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BACKGROUND: Activating and inhibitory KIR receptors (aKIR, iKIR) control the development and function of NK cells whose function alterations adjust the tumor microenvironment immunity. This research was conducted to determine the KIRs gene impact on genetic predisposition to Head and Neck Squamous Cell Carcinoma (HNSCC) in Iranians. METHODS: KIR genotyping using sequence-specific primers-polymerase chain reaction (SSP-PCR) method was performed to identify the presence of all 16 KIR genes in 285 HNSCC patients, including laryngeal, oral cavity and pharyngeal SCC and 273 controls (CNs). RESULTS: Comparison of KIRs gene frequency between HNSCC and CNs revealed a highly significant increase in KIR2DL5, 2DS1, 2DS5, 3DS1 and CxT4 genotype and a decrease in KIR2DS4 deleted variant and AA genotype carriers. A significant increase was noted in individuals withhigher iKIRs than aKIRs in HNSCC compared with CNs. Individuals with ≥4 iKIR and those with ≥5 aKIRs were significantly more common in HNSCC than CNs. 68distinct KIR genotypes were identified in 558 individuals. CONCLUSION: Our findings determined the detrimental impact of KIR2DS1, 2DS5, 3DS1, 2DL5 and CxT4 genotype as well as the protective impact of KIR2DS4del and AA genotype on genetic predisposition to HNSCC in Iranians.

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Carcinoma de Células Escamosas/genética , Neoplasias de Cabeza y Cuello/genética , Receptores KIR2DL5/genética , Receptores KIR3DS1/genética , Receptores KIR/genética , Anciano , Secuencia de Bases/genética , Carcinoma de Células Escamosas/epidemiología , Carcinoma de Células Escamosas/inmunología , Estudios de Casos y Controles , Femenino , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Neoplasias de Cabeza y Cuello/epidemiología , Neoplasias de Cabeza y Cuello/inmunología , Humanos , Irán/epidemiología , Células Asesinas Naturales/inmunología , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Receptores KIR/inmunología , Receptores KIR2DL5/inmunología , Receptores KIR3DS1/inmunología , Eliminación de Secuencia , Carcinoma de Células Escamosas de Cabeza y Cuello , Microambiente Tumoral/genética , Microambiente Tumoral/inmunología

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BACKGROUND: Febrile non-hemolytic transfusion reaction (FNHTR) is the most common type of transfusion reactions, and it could be reduced by transfusing patients with leukocyte-poor blood products. However, FNHTR still occur in certain patients transfused with leukocyte-poor red blood cell (LPR) products. It is examined whether human platelet antigen (HPA) could be a potential membrane antigen that plays a role in FNHTR. METHODS: A total of 120 inpatient subjects who transfused with LPR (60 in FNHTR group, 60 in control group) were typed for HPA-2, HPA-3, and HPA-15 using sequence specific primer-polymerase chain reaction (SSP-PCR) and electrophoresis. RESULTS: HPA-2 unmatched rate between donors and patients in FNHTR group was 18%, and only 3% unmatched rate was observed in control group (p=0.0082). FNHTR group was further classified according to the imputability. There was a significant difference (p=0.0041) between FNHTR (probable imputability, infection) group and control group, and more significant difference (p=0.0008) was seen between FNHTR (probable imputability, febrile neutropenia) group and control group. CONCLUSIONS: Those results indicated that HPA-2 might play roles on inducing FNHTR in patients suffering from infectious diseases and febrile neutropenia. HPA-2 genotyping between donors and recipients might be worth integrating in pre-transfusion testing to increase transfusion safety.

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Antígenos de Plaqueta Humana/genética , Fiebre/complicaciones , Reacción a la Transfusión/complicaciones , Reacción a la Transfusión/genética , Genotipo , Humanos

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We developed a sequence-specific primer PCR (SSP-PCR) for detection of a 5.8-kb deletion (B(m) 5.8) involving an erythroid cell-specific regulatory element in intron 1 of the ABO blood group gene. Using this SSP-PCR, we performed genetic analysis of 382 individuals with Bm or ABm. The 5.8-kb deletion was found in 380 individuals, and disruption of the GATA motif in the regulatory element was found in one individual. Furthermore, a novel 3.0-kb deletion involving the element (B(m) 3.0) was demonstrated in the remaining individual. Comparisons of single-nucleotide polymorphisms and microsatellites in intron 1 between B(m) 5.8 and B(m) 3.0 suggested that these deletions occurred independently.

Asunto(s)

Sistema del Grupo Sanguíneo ABO/genética , Células Eritroides/metabolismo , Eliminación de Gen , Intrones , Regiones Promotoras Genéticas , Humanos , Datos de Secuencia Molecular , Fenotipo , Polimorfismo de Nucleótido Simple

RESUMEN

BACKGROUND AND AIMS: The characterization of candidate gene polymorphisms in elderly populations is an important tool for the identification of risk factors for age-related diseases and conditions. We aimed to genotype the APOE polymorphisms (rs429358 and rs7412), rs61886492 (1561C>T) and rs202720 of GCPII gene and rs3918242 (-1562C>T) of MMP9 gene in an older-adult/elderly cohort from Cuiabá city, Mato Grosso Brazil as well as to characterize risk factors for morbidities and conditions affecting this cohort. METHODS: The studied population consisted of 570 subjects from Cuiabá city, Brazil, who were subjected to clinical interviews and blood collection for laboratory examinations and DNA extraction. Restriction Fragment Length Polymorphism Polymerase Chain Reaction (RFLP-PCR), sequence-specific primer PCR (SSP-PCR) and TaqMan® allelic discrimination assay were used for genotyping. RESULTS: The frequencies of APOE ε2 and ε4 were 6.6% and 14.8%, respectively, and the frequencies of GCPII rs61886492 T allele, GCPII rs202720 C allele and MMP9 rs3918242 T allele were, respectively, 3.0%, 26.6% and 10.1%. Significant associations between APOE ε2 allele with lower total cholesterol and LDL-cholesterol were found. In addition, MMP9 rs3918242 T allele was associated with higher LDL-cholesterol levels, suggesting a link between lipid metabolism alteration and cardiovascular disease. CONCLUSIONS: The present findings contributed to characterize risk factors specific for the studied population and to better understand the molecular physiopathology of common morbidities and conditions affecting older-adult/elderly people.

Asunto(s)

Apolipoproteínas E/genética , Carboxipeptidasas/genética , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Lípidos/sangre , Metaloproteinasa 9 de la Matriz/genética , Polimorfismo Genético , Factores de Edad , Anciano , Alelos , Brasil , Femenino , Genotipo , Humanos , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple

RESUMEN

Previous studies have shown weak associations between human dilated cardiomyopathy (DCM) and certain human leucocyte antigen (HLA) class II polymorphisms. Using a sequence-specific primer-PCR (SSP-PCR) technology, we compared the allelic distribution in the HLA-DQ and -DR locus in a cohort of German DCM patients (n=165) and DCM-free controls (n=79). With the exception of HLA-DQB1 0309, we found no significant differences between the two groups, even without adjustment for multiple testing. The HLA-DQB1 0309 allele, however, was detected more frequently in DCM patients as compared to controls (28.5% versus 10.1%, p=0.0010), leading to an odds ratio of 3.5 (95% confidence interval=1.5-9.1). The frequency of this allele was significantly higher in DCM patients without lymphocytic infiltrates in endomyocardial biopsies as compared to patients classified histologically as inflammatory DCM (33.1% versus 14.6%, p=0.028). There was no significant difference in the allelic HLA-DQB1 0309 distribution between DCM patients with and without viral genomes detected in the heart (24.2% versus 29.5%, p=0.668). In summary, the frequency of the HLA-DQB1 0309 allele is overrepresented in DCM patients, suggesting that carriers of this HLA class II variant are associated with an increased risk for developing DCM. Although Bonferroni adjustment was applied, controlled studies in larger samples of DCM patients and in different ethnic populations are warranted to confirm this observation and reveal the pathophysiological mechanisms behind this association.

Asunto(s)

Cardiomiopatía Dilatada/genética , Cadenas beta de HLA-DQ/genética , Adulto , Anciano , Alelos , Cardiomiopatía Dilatada/epidemiología , Estudios de Casos y Controles , Estudios de Cohortes , Femenino , Frecuencia de los Genes , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Alemania/epidemiología , Antígenos de Histocompatibilidad Clase II/genética , Humanos , Masculino , Persona de Mediana Edad

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Several host genetic factors play an important role in susceptibility to human immunodeficiency virus type 1 (HIV-1) infection and in its progression to acquired immune deficiency syndrome (AIDS). The interleukin-18 (IL-18) is a multifunctional proinflammatory cytokine that regulates immune responses and plays a pathogenic role in HIV-1 infection by enhancing viral replication. Single nucleotide polymorphisms (SNPs) in the IL-18 gene promoter region may lead to altered transcriptional activity and IL-18 production, and may account for variation in the risk of HIV-1 infection. We have investigated the association between IL-18 promoter polymorphism -607C>A and HIV-1 infection through a case-control study of 500 patients with HIV-1/AIDS and an equal number of age and sex matched controls in a north Indian population. Genotyping using sequence specific primer-polymerase chain reaction (SSP-PCR) showed a statistically significant reduced risk of HIV-1 infection for the A>A genotype [odds ratio (OR) = 0.57, 95% confidence interval (95% CI) = 0.33-0.98, p = 0.040], but not for the C>A genotype (OR = 0.87, 95% CI = 0.66-1.14, p = 0.321). We concluded that the -607A allele of the IL-18 gene promoter polymorphism may play a protective role against the progression of HIV-1 infection in this population.

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Objective To evaluate the practical value of Serology, PCR-SSP and PCR-SSOP for HLA-B typing. Methods A total of 30 samples, the blood of patients and donors waiting kidney transplantation, were used in the study. HLA-B typing was performed by one-step monoclonal antibody typing, micro PCR-SSP typing and PCR-SSOP reverse hybridisation. Results All samples were successfully typed by three methods. The difference between serological and PCR-SSP typing was 13%. The difference between PCR-SSOP and PCR-SSP typing was 3%. Conclusion Serology is high discrepancy rate and low-resolution, but cheap, simple and rapid. PCR-SSP and PCR-SSOP typing are high specific and accuracy. SSP is suitable for several samples, and SSOP is for lots of samples simultaneously although it needed long time.

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Objective:To establish a new method performed on an DNA chip for genotyping HLA DR and supply a new view.Methods:According to the particular sequence of HLA DR exon2,HLA DR genotyping Chip was made,then the labeled PCR products hybridized with them,the signals were sanned by sanner and analyzed by Imagene software.70 standard DNA and 200 donor recipients have been genotyped and some of samples have been sequenced.Results:The experimental results showed that the HLA DR genotyping chip made are accurate and sensitive.Thirty alleles of HLA DR were accurately distinguished and its overall time of DNA typing was 3 hours.Conclusion:This proved that the DNA Microarray technique is good for DR genotyping and high resolution,high specificity,well reproducibility.Compared with PCR SSP and PCR SSO methods,the genotyping chip method is more intuitionistic and has the advantage of integration.It can also genotype HLA A,B alleles and many persons in a chip at the same time.It is more suitable for clinical application and establishment of marrow bank and umbilical cord bank.