Han Min, Kang Xing, Liu Zhengbin, Zhang Tingting, Li Yanwei, Chen Chao, Wang Huijuan
The National Engineering Research Center for Miniaturized Detection Systems, College of Life Science, Northwest University , Xi'an, China .
AIDS Res Hum Retroviruses. 2017 Jul;33(7):711-717. doi: 10.1089/aid.2016.0280. Epub 2017 Mar 27.
HLA-B57:01 is strongly associated with severe adverse drug reaction induced by the anti-HIV drug abacavir (ABC) and antibiotic flucloxacillin. This study was dedicated to establishing a new method for HLA-B57:01 genotyping and investigating the HLA-B57:01 distribution pattern in four Chinese populations. A single-tube duplex real-time polymerase chain reaction (PCR) system was established by combining the amplification refractory mutation system and TaqMan probe. The reliability of this assay was validated by comparing the genotyping results with those by sequence-based typing. With this assay, the distribution of HLA-B57:01 in 354 blood samples from four ethnic groups, namely, Han, Tibetan, Uighur, and Buyei, was determined. A 100% concordance was observed between the results of real-time PCR and sequence-based typing in 50 Uighur samples. As low as 0.016 ng DNA that carried HLA-B57:01 could be detected with this assay. HLA-B57:01 carriers identified in 100 Northern Han Chinese, 104 Buyeis, 100 Tibetans, and 50 Uighurs were 0, 1 (0.96%), 3 (3%), and 6 (12%), respectively. The carrier rate of HLA-B57:01 in Uighur was significantly higher than those in Northern Han (p = .001) and Buyei (p = .005). The newly established real-time PCR assay provides a rapid and reliable tool for HLA-B57:01 allele screening before the prescription of ABC and flucloxacillin in clinical practice.
HLA - B57:01与抗HIV药物阿巴卡韦（ABC）和抗生素氟氯西林引起的严重药物不良反应密切相关。本研究致力于建立一种新的HLA - B57:01基因分型方法，并调查其在四个中国人群中的分布模式。通过结合扩增阻滞突变系统和TaqMan探针建立了单管双重实时聚合酶链反应（PCR）系统。通过将基因分型结果与基于序列分型的结果进行比较，验证了该检测方法的可靠性。利用该检测方法，确定了汉族、藏族、维吾尔族和布依族四个民族的354份血样中HLA - B57:01的分布情况。在50份维吾尔族样本中，实时PCR结果与基于序列分型的结果之间观察到100%的一致性。该检测方法能够检测到低至0.016 ng携带HLA - B57:01的DNA。在100名中国北方汉族、104名布依族、100名藏族和50名维吾尔族中鉴定出的HLA - B57:01携带者分别为0、1（0.96%）、3（3%）和6（12%）。维吾尔族中HLA - B57:01的携带率显著高于中国北方汉族（p = 0.001）和布依族（p = 0.005）。新建立的实时PCR检测方法为临床实践中在开具ABC和氟氯西林处方前进行HLA - B*57:01等位基因筛查提供了一种快速可靠的工具。
