CAJ | 학술논문

以铜绿微囊藻（Microcystis aeruginosa）16S rRNA基因片段为靶序列设计一对特异性引物，采用Real-time PCR法，对铜绿微囊藻进行定性、定量检测。试验表明，仅含铜绿微囊藻DNA模板的样品有特异性扩增，扩增产物熔解曲线平稳，峰尖且窄，熔解温度为（87±1）℃。以重组质粒pMD-18T-16S为标准品，检测区间为1．1×102 copies/mL～1．1×108 copies/mL，所得标准曲线符合制备实时定量PCR标准曲线的要求，对标准品进行测定，方法检出限为11 copies/mL。用该标准曲线对实验室培养获得的铜绿微囊藻DNA样品进行定量检测，与显微镜计数结果基本一致。
이동록미낭조（Microcystis aeruginosa）16S rRNA기인편단위파서렬설계일대특이성인물，채용Real-time PCR법，대동록미낭조진행정성、정량검측。시험표명，부함동록미낭조DNA모판적양품유특이성확증，확증산물용해곡선평은，봉첨차착，용해온도위（87±1）℃。이중조질립pMD-18T-16S위표준품，검측구간위1．1×102 copies/mL～1．1×108 copies/mL，소득표준곡선부합제비실시정량PCR표준곡선적요구，대표준품진행측정，방법검출한위11 copies/mL。용해표준곡선대실험실배양획득적동록미낭조DNA양품진행정량검측，여현미경계수결과기본일치。
A real-time polymerase chain reaction ( PCR) assay was designed and evaluated for rapid detec-tion and quantification of the algae Microcystis aeruginosa.A pair of specific primers was designed from the se-quence of 16S region,of which the PCR pecificity was examined compared with Chlorella sp.and Nitzschia sp.. PCR amplifications were detected only from samples which contained Microcystis cells and specific signals were not detected from Chlorella sp..Melting curve was stationary and peak was narrow .Melting temperature was (87 ±1)℃.Based on recombinant plasmid pMD -18T-16S for the standard,the standard curve we got has high linearity and correlation coeficient .Moreover,this assay was in accordance with preparative real-time PCR. Using the developed standard curves ,Microcystis aeruginosa could be quantified in agreement with the quantifica-tion by optical microscopy .