Center for Developmental Therapeutics, Seattle Children's Research Institute, Seattle, Washington, USA.
Genet Med. 2011 Sep;13(9):794-9. doi: 10.1097/GIM.0b013e31821afca5.
The diagnosis of a mitochondrial disorder relies heavily on the enzymatic analysis of mitochondrial respiratory chain complexes in muscle or other tissues. However, considerable differences exist between clinical laboratories in the protocols or particular tests used for evaluation. In addition, laboratories can encounter difficulties in consistent technique, as well as procurement of adequate positive or negative controls. Currently, there is no external quality assurance for respiratory chain complex assays. In this study, we explored the use of Caenorhabditis elegans mitochondria as a potential aid to diagnostic centers that perform respiratory chain complex assays.
Five diagnostic test centers in the United States and one from Australia comparatively analyzed enzyme activities of mitochondria from C. elegans. The first survey consisted of three open-labeled samples including one normal control and two mutants; the second survey consisted of one open-labeled normal control and two blinded samples.
There was very good concordance among laboratories in detecting the majority of the defects present in the mutant specimens. Despite the ability to detect respiratory chain complex defects, the scatter between centers for certain enzymatic assays, particularly I + III, II, III, and IV, led to different diagnostic interpretations between the centers.
The data strongly support the need for comparative testing of mitochondrial enzyme assays between multiple laboratories. Our overall results are encouraging for the use of nematode mitochondria as a tool that might provide a virtually inexhaustible supply of mitochondria with defined defects for development of assays and as a potential source of control specimens.
线粒体疾病的诊断主要依赖于肌肉或其他组织中线粒体呼吸链复合物的酶分析。然而，不同临床实验室在评估中使用的方案或特定检测方法存在很大差异。此外，实验室在技术的一致性以及获得充足的阳性或阴性对照方面可能会遇到困难。目前，呼吸链复合物检测没有外部质量保证。在这项研究中，我们探讨了利用秀丽隐杆线虫线粒体作为为进行呼吸链复合物检测的诊断中心提供潜在帮助的方法。
美国的五个诊断测试中心和一个来自澳大利亚的中心比较分析了秀丽隐杆线虫线粒体的酶活性。第一次调查包括三个开放标签样本，包括一个正常对照和两个突变体；第二次调查包括一个开放标签的正常对照和两个盲样。
尽管能够检测到突变体样本中存在的大多数缺陷，但各实验室在检测大多数缺陷方面非常一致。尽管能够检测到呼吸链复合物缺陷，但某些酶检测，特别是 I + III、II、III 和 IV 之间的中心散射，导致中心之间的诊断解释不同。
数据强烈支持需要在多个实验室之间进行线粒体酶检测的比较测试。我们的总体结果令人鼓舞，因为线虫线粒体可用作工具，为开发检测方法提供几乎用之不竭的具有明确缺陷的线粒体，并作为对照样本的潜在来源。
