High-resolution HLA-DQB1 typing using the polymerase chain reaction and sequence-specific primers

Polymorphism at HLA-DQB1 is known to influence tissue compatibility and disease susceptibility; however, current DQB1 typing methods are unable to distinguish the 32 currently recognized DQB1 alleles. We have developed a 32-reaction PCR-SSP method capable of differentiating all DQB1 alleles that differ in amino acid sequence. This method can resolve all heterozygous combinations of DQB1 alleles, with the exception of several combinations involving alleles not thus far detected in Caucasoid populations.     

Determination of Lewis FUT3 gene mutations by PCR using sequence‐specific primers enables efficient genotyping of clinical samples

We have developed a polymerase chain reaction method using sequence‐specific primers (PCR‐SSP) for rapid and correct genotyping of the common Lewis (FUT3) gene mutations 59T>G, 202T>C, 314C>T, 508G>A, and 1067T>A. The PCR‐SSP method was validated on 20 healthy blood donors and 16 non‐insulin‐dependent diabetic patients. All individuals were in parallel genotyped by our established polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) analysis. The FUT3 genotypes, determined with the PCR‐SSP method, were in complete accordance with the results of the PCR‐RFLP reference method. The PCR‐SSP method could also be adapted to assign the presence of a specific mutation to the respective FUT3 alleles. We found the method to be reliable, rapid and cheap with no requirements for restriction enzyme processing. Hum Mutat 18:358–359, 2001. © 2001 Wiley‐Liss, Inc.     

The HLA-B*15:02 polymorphism and Tegretol®-induced serious cutaneous reactions in epilepsy: An updated systematic review and meta-analysis

Tegretol^® [carbamazepine (CBZ)], an aromatic drug approved for epilepsy treatment, can induce adverse drug reactions (ADRs) after its administration. Several genetic studies of epilepsy have shown that genetic polymorphisms increase the risk of ADRs, and some interactions between CBZ and other treatments can also induce adverse effects. Thus, to avoid such interactions and to provide an overview of the genetic profiles involved in ADRs with CBZ, for the first time, a systematic review and meta-analysis focusing on epilepsy was performed, using Cochrane Library, Embase and PubMed databases to find studies published between January 1980 and October 2016. Of the eligible studies, those selected were related to the impact of genetic polymorphisms on ADRs in patients receiving antiepileptic treatment. The results of these selected studies are expressed as pooled odds ratios (ORs) with 95% confidence intervals (CIs), based on data from individual patients. Out of 807 articles, nine were included in the present meta-analysis to assess the association between human leukocyte antigen (HLA)-B*15:02 polymorphisms and CBZ-induced serious cutaneous reactions (SCRs), such as Stevens–Johnson syndrome (SJS) and toxic epidermal necrolysis (TEN), in epilepsy. It was found that HLA-B*15:02 polymorphisms were significantly associated with CBZ SCR risk (OR: 27.325, 95% CI: 9.933–51.166), while subgroup analyses by ethnicity showed that the association was significant in Han Chinese (OR: 42.059, 95% CI: 9.587–184.514). The HLA-B*15:02 polymorphism was also strongly associated with the CBZ-SJS subgroup (OR: 152.089, 95% CI: 34.737–665.901) and significantly associated with the CBZ-SJS/TEN subgroup (OR: 13.993, 95% CI: 7.291–26.856). Also, the allele was overrepresented in the Han Chinese population (OR: 17.886, 95% CI: 8.411–38.034) within the CBZ-SJS/TEN subgroup. Although the number of studies available in other Asian ethnicities was insufficient for determining publication bias, it nevertheless showed a relationship between the HLA-B*15:02 polymorphism and SCRs. In addition, despite the small number of included studies, the results reveal strong evidence that the HLA-B*15:02 polymorphism can induce SCRs among Asian CBZ users. These findings should prompt physicians to individualize CBZ therapy for patients with epilepsy.     

螺旋CT性能模体和扫描轮廓线检测初步探讨

对螺旋CT的检测模体及检测方法进行了调研试用,并对螺旋CT扫描轮廓线(SSP)进行了初步测试与分析.     

HLA-DRB1基因分型在移植配型及亲子鉴定中的应用

应用聚合酶链反应－序列持异性引物（ＰＣＲ＿ＳＳＰ）方法对南京地区汉族人群ＨＬＡ－ＤＲＢ１位点基因特异性及骨髓移植配型、亲子鉴定等进行了分析。ＤＲＢ１基因频率范围在０．００３３～０．１８３３之间，以ＤＲ９１ＤＲ４占多数；骨髓移植通过配型移植成功３例，２例存活情况良好；ＤＲＢ１位点排除亲子关系５例，其中２例为ＨＬＡ－ＤＲＢ１单独排除亲缘关系。本法具有操作简单、快速、结果可靠的特点，不仅适用于移植配型、     

福建汉族系统性红斑狼疮与HLA-DR基因的关联研究

目的 探讨HLA－DR多态性与SLE的易感性关系。方法 用聚合酶链反应－序列特异引物（PCR－SSP）方法,对80例SLE患者和104例健康人HLA－DR基因进行研究。结果 SLE患者DR15基因频率明显升高（P＜0．001,RR＝3．86）,DR3、DR4基因频率显著下降（P＜0．05,RR＝0．48;P＜0．05,RR0．38）,DR15阳性的患者中狼疮性肾炎发病率高于其它患者（P＜0．05,RR＝3．15）,抗SSA抗体也明显升高（P＜0．05,RR＝3．46）。     

一个新的等位基因:HLA-A~*1114克隆和序列分析

目的　识别确认中国人群的HLA新等位基因。方法　使用PCR SSP以及以测序为基础的分型 (SBT)技术 ,在HLA分型常规工作中发现 1个与HLA A 110 2基因相关的新基因 ,以基因克隆及DNA测序 ,分析该基因和A 110 2基因序列的差异。结果　该基因和A 110 2基因序列的差异在于外显子 3区域中 ,5 2 4A >G ,5 2 6G>C以及 5 2 7C >G等 3个碱基的取代 ,使密码子 175由组氨酸变为精氨酸 ,密码子 176由丙氨酸变为精氨酸。结论该基因为HLA新等位基因 ,2 0 0 2年 9月已被世界卫生组织HLA因子命名委员会正式命名为HLA A 1114。用PCR SSP方法 ,在 30 0 0名随机造血干细胞供者中 ,未发现其他带有A 1114基因的个体。