Park Tae-Gyu, de Salas Miguel F, Bolch Christopher J S, Hallegraeff Gustaaf M
School of Plant Science, University of Tasmania, Private Bag 55, Churchill Avenue, Sandy Bay, Hobart, Tasmania, Australia.
Appl Environ Microbiol. 2007 Apr;73(8):2552-60. doi: 10.1128/AEM.02389-06. Epub 2007 Feb 23.
A TaqMan format real-time PCR probe was developed against the internal transcribed spacer 2 ribosomal DNA region for the specific detection and quantification of Cryptoperidiniopsis brodyi in environmental samples. The assay specificity was confirmed by testing against related dinoflagellates and verified by sequencing PCR amplicons from natural water samples. Phylogenetic analysis of the sequenced environmental samples also showed that this assay is specific to C. brodyi. The C. brodyi-specific assay was used in conjunction with Pfiesteria piscicida- and Pfiesteria shumwayae-specific real-time PCR assays to investigate the temporal variations of C. brodyi, P. piscicida, and P. shumwayae abundance in the Derwent estuary, Tasmania. The 18-month field survey from November 2004 to April 2006 revealed that C. brodyi occurred in all seasons at very low densities, mostly below 25 cells liter(-1), with higher abundance (maximum, 112 cells liter(-1)) in April and May. P. piscicida was detected only once, in May 2005 at 60 cells liter(-1). P. shumwayae was not detected during the survey.
针对核糖体DNA内转录间隔区2开发了一种TaqMan格式实时荧光定量PCR探针，用于环境样品中布氏隐多甲藻（Cryptoperidiniopsis brodyi）的特异性检测和定量分析。通过与相关甲藻进行检测来确认该检测方法的特异性，并通过对天然水样的PCR扩增产物进行测序来验证。对测序后的环境样品进行系统发育分析也表明该检测方法对布氏隐多甲藻具有特异性。将布氏隐多甲藻特异性检测方法与杀鱼费氏藻（Pfiesteria piscicida）和舒氏费氏藻（Pfiesteria shumwayae）特异性实时荧光定量PCR检测方法结合使用，以研究塔斯马尼亚德文特河口布氏隐多甲藻、杀鱼费氏藻和舒氏费氏藻丰度的时间变化。2004年11月至2006年4月为期18个月的实地调查显示，布氏隐多甲藻在所有季节均以极低密度出现，大多低于25个细胞/升，在4月和5月丰度较高（最高达112个细胞/升）。杀鱼费氏藻仅在2005年5月被检测到一次，为60个细胞/升。在调查期间未检测到舒氏费氏藻。
