Bai Shumeng, Tian Li, Shi Zhengping, Liu Ning, Zhang Jiuming
First Institute of Oceanography, State Oceanic Administration, Qingdao 266061, China.
Wei Sheng Wu Xue Bao. 2010 Jan;50(1):76-80.
PCR method was used to detect Penicillium griseofulvum, a dominant species in marine contaminated sediments and thereby to deduce the contamination degree.
According to differences in internal transcribed space (ITS) sequences of Penicillium genus and specific IAO sequence, we designed species-specific primers AS1/RS4 and IAO1/IAO2 of Penicillium griseofulvum and established the corresponding PCR systems. By using PCR and nested-PCR, the detection sensitivity was compared.
The primers could exclusively amplify destined DNA fragment from environment. Using AS1/RS4 as primers, the detection sensitivity could be 10 fg/microL and 10 spores. The detection sensitivity for the sediments was 10(2) spores/0.25 g sediments. While the detection was unsensitive when using IAO1/IAO2 as primers.
It is feasible that the species-specific primers be used as probes for the detection of environmental pollution dominant species, Penicillium griseofulvum, because the frequency of occurrence and amount of this strain could preferably indicate the pollution degree.
采用PCR方法检测海洋污染沉积物中的优势种灰黄青霉，从而推断污染程度。
根据青霉属内转录间隔区（ITS）序列差异及特定IAO序列，设计灰黄青霉的种特异性引物AS1/RS4和IAO1/IAO2，并建立相应的PCR体系。通过PCR和巢式PCR比较检测灵敏度。
引物能特异性地从环境中扩增出目的DNA片段。以AS1/RS4为引物，检测灵敏度可达10 fg/μL和10个孢子。沉积物的检测灵敏度为10²个孢子/0.25 g沉积物。而以IAO1/IAO2为引物时检测不灵敏。
种特异性引物作为检测环境污染优势种灰黄青霉的探针是可行的，因为该菌株的出现频率和数量能较好地指示污染程度。
