Keer J T, Birch L
BioAnalytical Innovation Team, LGC Limited, Queens Road, Teddington, Middlesex TW11 0LY, UK.
J Microbiol Methods. 2003 May;53(2):175-83. doi: 10.1016/s0167-7012(03)00025-3.
A significant number of pathogenic microorganisms can be found in environmental reservoirs (air, water, soil). It is important to assess the viability status of these organisms to determine whether they pose a threat to public health. Classical methods for determining viability are time consuming. Hence, molecular methods have been developed to address this problem. Molecular methods offer speed, sensitivity and specificity. Both DNA and RNA have been analysed using molecular amplification methods such as polymerase chain reaction (PCR), reverse transcriptase PCR (RT-PCR) and nucleic acid sequence-based amplification (NASBA). However, due to the variable persistence of nucleic acids in cells post-death, the correlation between presence of DNA and RNA and viability is not clear-cut. Similarly, the choice of target and sensitivity of the method can significantly affect the validity of the viability assay. This review assesses the molecular methods currently available and evaluates their ability to assess cell viability with emphasis on environmental pathogens.
大量致病微生物可在环境储存库（空气、水、土壤）中被发现。评估这些微生物的生存状态对于确定它们是否对公众健康构成威胁很重要。传统的确定生存能力的方法耗时较长。因此，已开发出分子方法来解决这一问题。分子方法具有速度快、灵敏度高和特异性强的特点。DNA和RNA都已通过诸如聚合酶链反应（PCR）、逆转录PCR（RT-PCR）和基于核酸序列的扩增（NASBA）等分子扩增方法进行分析。然而，由于细胞死亡后核酸存在不同程度的持续性，DNA和RNA的存在与生存能力之间的相关性并不明确。同样，方法的靶标选择和灵敏度会显著影响生存能力测定的有效性。本综述评估了目前可用的分子方法，并着重针对环境病原体评估了它们评估细胞生存能力的能力。
