Burlinson Brian
Safety Assessment, Huntingdon Life Sciences, Woolley Road, Alconbury Huntingdon, Cambridgeshire, PE28 4HS, UK.
Methods Mol Biol. 2012;817:143-63. doi: 10.1007/978-1-61779-421-6_8.
The strategy for testing for genotoxicity covers three main areas, namely gene mutation, chromosome aberration or breakage (clastogenicity), and chromosome loss or gain (aneuploidy). The current generalized strategy consists of assays capable of detecting all of these endpoints using in vitro assays such as the Ames test for detecting gene mutations in bacteria, the human peripheral lymphocyte chromosome aberration (CA) test for detecting clastogenicity, and the in vitro micronucleus test for clastogenicity and aneuploidy. The primary in vivo assay, and generally the only in vivo assay required, is the in vivo rodent bone marrow micronucleus assay. However, there are instances when these assays alone are inadequate and further testing is required, especially in vivo. Historically, the preferred second assay has been the rodent liver unscheduled DNA synthesis assay but recently this has been superseded by the rodent single cell gel electrophoresis or Comet assay. This assay has numerous advantages especially in vivo, where virtually any tissue can be examined. The status of the in vitro comet assay in regulatory testing is much less clear although a preliminary review of data from the assay has shown it to be more specific than other in vitro genotoxicity tests and less prone to false positives.Detailed here are general protocols for both the in vitro and in vivo comet assays which will form the basis of the pending OECD guideline for the assay.
遗传毒性检测策略涵盖三个主要领域，即基因突变、染色体畸变或断裂（致断裂性）以及染色体丢失或增加（非整倍体）。当前的通用策略包括一系列检测方法，这些方法能够通过体外试验检测所有这些终点，如用于检测细菌基因突变的艾姆斯试验、用于检测致断裂性的人类外周血淋巴细胞染色体畸变（CA）试验以及用于检测致断裂性和非整倍体的体外微核试验。主要的体内试验，通常也是唯一需要的体内试验，是体内啮齿动物骨髓微核试验。然而，在某些情况下，仅这些试验是不够的，需要进一步检测，尤其是体内检测。从历史上看，首选的第二种试验是啮齿动物肝脏非程序性DNA合成试验，但最近它已被啮齿动物单细胞凝胶电泳或彗星试验所取代。该试验有许多优点，尤其是在体内，几乎可以检查任何组织。体外彗星试验在监管检测中的地位尚不太明确，尽管对该试验数据的初步审查表明它比其他体外遗传毒性试验更具特异性，且不易出现假阳性。这里详细介绍了体外和体内彗星试验的一般方案，这些方案将构成即将出台的经合组织该试验指南的基础。
