Hogan Kirk J, Burmester James K, Caldwell Michael D, Hogan Quinn H, Coursin Douglas B, Green Dawn N, Selzer Rebecca M R, Broderick Thomas P, Rusy Deborah A, Poroli Mark, Lutz Anna L, Sanders Allison M, Oldenburg Mary C, Koelbl James A, de Arruda-Indig Monika, Halsey Jennifer L, Day Stephen P, Domanico Michael J
Department of Anesthesiology, University of Wisconsin School of Medicine and Public Health, B6/319 Clinical Sciences Center, 600 Highland Avenue, Madison, WI 53792, USA.
Clin Med Res. 2009 Sep;7(3):69-84. doi: 10.3121/cmr.2009.837. Epub 2009 May 27.
Many complications in the perioperative interval are associated with genetic susceptibilities that may be unknown in advance of surgery and anesthesia, including drug toxicity and inefficacy, thrombosis, prolonged neuromuscular blockade, organ failure and sepsis. The aims of this study were to design and validate the first genetic testing platform and panel designed for use in perioperative care, to establish allele frequencies in a target population, and to determine the number of mutant alleles per patient undergoing surgery. DESIGN/SETTING/PARTICIPANTS AND METHODS: One hundred fifty patients at Marshfield Clinic, Marshfield, Wisconsin, 100 patients at the Medical College of Wisconsin Zablocki Veteran's Administration Medical Center, Milwaukee, Wisconsin, and 200 patients at the University of Wisconsin Hospitals and Clinics, Madison, Wisconsin undergoing surgery and anesthesia were tested for 48 polymorphisms in 22 genes including ABC, BChE, ACE, CYP2C9, CYP2C19, CYP2D6, CYP3A4, CYP3A5, beta2AR, TPMT, F2, F5, F7, MTHFR, TNFalpha, TNFbeta, CCR5, ApoE, HBB, MYH7, ABO and Gender (PRKY, PFKFB1). Using structure-specific cleavage of oligonucleotide probes (Invader, Third Wave Technologies, Inc., Madison, WI), 96-well plates were configured so that each well contained reagents for detection of both the wild type and mutant alleles at each locus.
There were 21,600 genotypes confirmed in duplicate. After withdrawal of polymorphisms in non-pathogenic genes (i.e., the ABO blood group and gender-specific alleles), 376 of 450 patients were found to be homozygous for mutant alleles at one or more loci. Modes of two mutant homozygous loci and 10 mutant alleles in aggregate (i.e., the sum of homozygous and heterozygous mutant polymorphisms) were observed per patient.
Significant genetic heterogeneity that may not be accounted for by taking a family medical history, or by obtaining routine laboratory test results, is present in most patients presenting for surgery and may be detected using a newly developed genotyping platform.
围手术期的许多并发症与遗传易感性相关，这些遗传易感性在手术和麻醉前可能是未知的，包括药物毒性和无效、血栓形成、神经肌肉阻滞延长、器官衰竭和败血症。本研究的目的是设计并验证首个用于围手术期护理的基因检测平台和检测组，确定目标人群中的等位基因频率，并确定每位接受手术患者的突变等位基因数量。
设计/地点/参与者和方法：在威斯康星州马什菲尔德市马什菲尔德诊所的150例患者、威斯康星州密尔沃基市威斯康星医学院扎布洛斯基退伍军人管理局医疗中心的100例患者以及威斯康星大学医院及诊所（位于威斯康星州麦迪逊市）的200例接受手术和麻醉的患者，检测了22个基因中的48个多态性，这些基因包括ABC、BChE、ACE、CYP2C9、CYP2C19、CYP2D6、CYP3A4、CYP3A5、β2AR、TPMT、F2、F5、F7、MTHFR、TNFα、TNFβ、CCR5、ApoE、HBB、MYH7、ABO以及性别（PRKY、PFKFB1）。使用寡核苷酸探针的结构特异性切割（Invader，Third Wave Technologies公司，威斯康星州麦迪逊市），配置96孔板，使每个孔都含有用于检测每个位点野生型和突变等位基因的试剂。
共重复确认了21,600种基因型。去除非致病基因中的多态性（即ABO血型和性别特异性等位基因）后，发现450例患者中有376例在一个或多个位点上为突变等位基因纯合子。每位患者观察到两个突变纯合子位点的模式以及总共10个突变等位基因（即纯合和杂合突变多态性的总和）。
大多数接受手术的患者存在显著的遗传异质性，这可能无法通过家族病史或常规实验室检查结果来解释，并且可以使用新开发的基因分型平台进行检测。
