Makowski Gregory S, Nadeau Francesca L, Hopfer Sidney M
Department of Laboratory Medicine, University of Connecticut Health Center, Farmington, Connecticut 06030-2235, USA.
Ann Clin Lab Sci. 2003 Summer;33(3):243-50.
In response to recommendations for cystic fibrosis (CF) carrier screening of the American College of Medical Genetics/American College of Obstetrics and Gynecology (ACMG/ACOG), we evaluated a commercial kit for mutation panel testing (Roche CF Gold Linear Array Panel). This kit tests for 25 CF mutations and 4 polymorphisms; it comprises an analyte specific reagent for single tube multiplex polymerase chain reaction (PCR) amplification and subsequent allele specific oligonucleotide (ASO) hybridization. Neonatal whole blood samples collected on Guthrie card filter paper served as the DNA source. Following a wash step to remove whole blood PCR inhibitors, multiplex PCR amplification could be performed either on DNA that was heat extracted from Guthrie cards or directly on the filter paper matrix itself. In 13 CF patient samples, the CF mutation results obtained with this kit agreed completely with those obtained from a reference laboratory that performs an 87 CF mutation panel. The kit was reliable, despite the small sample size (3 mm diameter punch of the Guthrie card), the presence of PCR inhibitors in whole blood, and protracted storage of blood samples (up to 9 yr at room temperature). The kit was convenient, cost competitive, and practical for use in a small CF screening laboratory.
为响应美国医学遗传学学会/美国妇产科医师学会（ACMG/ACOG）关于囊性纤维化（CF）携带者筛查的建议，我们评估了一种用于突变检测的商业试剂盒（罗氏CF金线性阵列试剂盒）。该试剂盒可检测25种CF突变和4种多态性；它包含一种分析物特异性试剂，用于单管多重聚合酶链反应（PCR）扩增及随后的等位基因特异性寡核苷酸（ASO）杂交。采集在格思里卡片滤纸上的新生儿全血样本用作DNA来源。经过一个去除全血PCR抑制剂的洗涤步骤后，多重PCR扩增既可以对从格思里卡片上热提取的DNA进行，也可以直接在滤纸基质本身上进行。在13份CF患者样本中，该试剂盒获得的CF突变结果与一个进行87种CF突变检测的参考实验室所获得的结果完全一致。尽管样本量小（格思里卡片直径3毫米的冲孔样本）、全血中存在PCR抑制剂且血样储存时间长（室温下长达9年），该试剂盒依然可靠。该试剂盒使用方便、具有成本竞争力，适用于小型CF筛查实验室。
