CAJ | 학술논문

为建立快速、准确的赤潮藻鉴定和定量检测方法,以红色裸甲藻(Gymnodinium sanguineum)作为研究对象,将快速检测手段一实时荧光定量PCR(RFQ-PCR)技术应用于赤潮藻的种类鉴定中.首先以红色裸甲藻18S rDNA序列为靶区域.设计出适合用于RFQ-PCR技术的红色裸甲藻引物和TaqMan探针,并通过引物PCR验证确定其种特异性;进而应用实时荧光定量PCR方法建立红色裸甲藻快速检测方法.与显微镜计数方法比较,两者所获结果无显著性差异,且该方法的检测限远低于红色裸甲藻爆发赤潮时的藻密度,因此该方法可有效地应用于红色裸甲藻的检测,从而为红色裸甲藻赤潮的预警预测奠定基础.
위건립쾌속、준학적적조조감정화정량검측방법,이홍색라갑조(Gymnodinium sanguineum)작위연구대상,장쾌속검측수단일실시형광정량PCR(RFQ-PCR)기술응용우적조조적충류감정중.수선이홍색라갑조18S rDNA서렬위파구역.설계출괄합용우RFQ-PCR기술적홍색라갑조인물화TaqMan탐침,병통과인물PCR험증학정기충특이성;진이응용실시형광정량PCR방법건립홍색라갑조쾌속검측방법.여현미경계수방법비교,량자소획결과무현저성차이,차해방법적검측한원저우홍색라갑조폭발적조시적조밀도,인차해방법가유효지응용우홍색라갑조적검측,종이위홍색라갑조적조적예경예측전정기출.
For detecting the harmful algal bloom(HAB)species sensitively and rapidly,Gymnodinium sanguineum was taken as the object,and the rapid detection method-RFQ-PCR(real-time fluorescent quantitative polymerase chain reaction)technique was applied to the HAB species detection.Firstly,gene specific primers and a TaqMan DNA probe were designed based on the sequence of 18S rDNA cloned from G.sanguineum.Then,using the primers and probe combination.a RFQ-PCR method was developed for quantitative detection of G.sanguineum.a harmful algae bloom species.The RFQ-PCR assay data showed good agreement with traditional microscope counting methods,suggesting that RFQPCR may be a useful method for detecting G.sanguineum.The detection threshold tested was much lower than that of G.sanguineum HAB.so this technique will be useful for forecasting and preventing G.sanguineum HAB.