Martin Rosalind, Latten Mark, Hart Padraig, Murray Helena, Bailie Deborah A, Crockard Martin, Lamont John, Fitzgerald Peter, Graham Colin A
Northern Ireland Regional Genetics Centre, Belfast City Hospital, Belfast Health and Social Care Trust, Belfast, Northern Ireland, UK.
Randox Laboratories Ltd., Crumlin, Northern Ireland, UK.
Atherosclerosis. 2016 Nov;254:8-13. doi: 10.1016/j.atherosclerosis.2016.09.061. Epub 2016 Sep 17.
Familial hypercholesterolaemia (FH) leads to a lifelong increase in plasma LDL levels with subsequent increase in premature vascular disease. Early diagnosis and treatment is the key to effective management of this condition. This research aims to produce a simple and cost effective genetic test which could identify the majority (71%) of mutations causing FH in the UK and Ireland.
The Randox Biochip Array Technology was used to detect 40 point mutations in LDLR, APOB and PCSK9 genes, over two 5 × 5 arrays. This technology uses multiplex allele specific PCR and biochip array hybridisation, followed by a chemiluminescence detection system and software for automated mutation calling.
The FH biochip array assay was validated in the Belfast Genetics Laboratory using 199 cascade screening samples previously sequenced for known FH causing family mutations, the overall sensitivity was 98%. The assay was then used for routine testing of 663 patients with possible FH, from clinics across the UK and Ireland. A total of 49 (7.4%) mutation positive individuals were identified, however, for the clinics in England the detection rate was 12.9%. Further analysis of 120 biochip negative patients, using DNA sequencing, did not identify any false negatives.
The FH biochip array provides a rapid and reliable genetic test for the majority of FH causing point mutations in the UK and Ireland. A total of 32 samples can be run in 3 h. This allows clinics to evaluate additional patients for a possible diagnosis of FH such as patients with high LDL, patients with early onset coronary disease, and patients with relatives known to have FH.
家族性高胆固醇血症（FH）会导致血浆低密度脂蛋白（LDL）水平终生升高，进而增加早发性血管疾病的风险。早期诊断和治疗是有效管理该疾病的关键。本研究旨在开发一种简单且经济高效的基因检测方法，以识别在英国和爱尔兰导致FH的大多数（71%）突变。
使用兰多克斯生物芯片阵列技术在两个5×5阵列上检测LDLR、APOB和PCSK9基因中的40个点突变。该技术采用多重等位基因特异性PCR和生物芯片阵列杂交，随后是化学发光检测系统和用于自动突变检测的软件。
FH生物芯片阵列检测方法在贝尔法斯特遗传学实验室使用199个先前已测序以确定已知FH致病家族突变的级联筛查样本进行了验证，总体灵敏度为98%。然后该检测方法用于对来自英国和爱尔兰各地诊所的663例疑似FH患者进行常规检测。共鉴定出49例（7.4%）突变阳性个体，然而，对于英格兰的诊所，检测率为12.9%。对120例生物芯片检测阴性的患者进行DNA测序进一步分析，未发现任何假阴性结果。
FH生物芯片阵列为英国和爱尔兰大多数导致FH的点突变提供了快速可靠的基因检测。3小时内可运行32个样本。这使得诊所能够评估更多可能诊断为FH的患者，如LDL水平高的患者、早发性冠心病患者以及已知有FH亲属的患者。
