Application of real-time PCR assay for detection and quantification of bloom-forming diatom Chaetoceros tenuissimus Meunier
Application of real-time PCR assay for detection and quantification of bloom-forming diatom Chaetoceros tenuissimus Meunier
复制标题
应用实时荧光定量PCR检测和定量开花硅藻Chaetoceros tenuissimus Meunier
DOI：
10.3800/pbr.5.56
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发表时间：
2010
影响因子：
0.6
通讯作者：
Y. Tomaru
中科院分区：
文献类型：
--
作者：
Kensuke Toyoda;Keizo Nagasaki;Y. Tomaru
2008年发现了水华硅藻细角毛藻（Chaetoceros tenuissimus）的病毒感染，现在认为它对C。自然环境中的tenuissimus种群。C的枚举在原位检查宿主-病毒关系时，细薄壳藻及其病毒是必不可少的;然而，硅藻物种的尺寸如此之小，以至于几乎不可能通过光学显微镜对其进行识别和计数。为了解决这一问题，我们建立了一种基于TaqMan的实时聚合酶链反应（PCR）方法来检测和定量C。极细针对28 S核糖体RNA（rRNA）基因D1区设计引物和TaqMan探针，建立的实时荧光定量PCR方法在种水平上具有特异性。极细Tris-EDTA缓冲液煮沸法是一种有效提取滤捕C.薄肌细胞在这项研究中。建立的实时荧光定量PCR方法的检测范围为100 μ g/ml。tenuissimus为10 1 ~ 10 6个细胞，该方法适用于C.极细的
Viral infection of the bloom-forming diatom Chaetoceros tenuissimus was discovered in 2008, and is now assumed to have a significant influence on the dynamics of C. tenuissimus populations in natural environments. Enumeration of C. tenuissimus and its viruses is essential when examining the host-virus relationship in situ; however, the diatom species is so small in size that its identification and counting by optical microscope is almost impossible. To resolve this problem, we have developed a TaqMan-based real-time polymerase chain reaction (PCR) method for detec- tion and quantification of C. tenuissimus. We designed primers and a TaqMan probe to target the D1 region of its 28S ribosomal RNA (rRNA) gene; the established real-time PCR was specific at the species level by testing 41 microalgal strains including C. tenuissimus. Tris-EDTA buffer-based boiling method was shown to be efficient for extracting DNA from filter-trapped C. tenuissimus cells in this study. The detection range of the established TaqMan-based real-time PCR method for C. tenuissimus was 10 1 to 10 6 cells collected on a filter; the method was applicable for C. tenuissimus
影响因子：
2.2
作者：
Ryoma Kamikawa;J. Asai;T. Miyahara;Keisuke Murata;K. Oyama;S. Yoshimatsu;Takashi Yoshida;Y. Sako
通讯作者：
Ryoma Kamikawa;J. Asai;T. Miyahara;Keisuke Murata;K. Oyama;S. Yoshimatsu;Takashi Yoshida;Y. Sako
影响因子：
6.6
作者：
Kamikawa, Ryoma;Nagai, Satoshi;Sako, Yoshihiko
通讯作者：
Sako, Yoshihiko
影响因子：
4.4
作者：
Shirai, Yoko;Tomaru, Yuji;Nagasaki, Keizo
通讯作者：
Nagasaki, Keizo