Detection of Phaeocystis globosa using sandwich hybridization integrated with nuclease protection assay (NPA-SH).
Phaeocystis globosa Scherffel is one of the common harmful algae species in coastal waters of the southeastern China. In this study, sandwich hybridization integrated with nuclease protection assay (NPA-SH) was used to qualitatively and quantitatively detect P. globosa. Results showed that this method had good applicability and validity in analyzing the samples from laboratory cultures and from fields. The linear regression equation for P. globosa was obtained, and the lowest detection number of cells was 1.8 x 10(4) cells. Statistics showed that there was no distinct difference between the results of detecting the microalgae by NPA-SH and traditional microscopy. This technique has good reliability, accuracy, and can give a remarkably high sample processing rate. Sandwich hybridization integrated with nuclease protection assay will provide an efficient alternative to microscopic method for monitoring and investigating the bloom of P. globosa.
Zhen
Yu
Y
Key Laboratory of Marine Environment and Ecology, Ministry of Education, Ocean University of China, Qingdao 266100, China. zhenyu@ouc.edu.cn
Mi
Tiezhu
T
Yu
Zhigang
Z
D002138:Calibration; D056890:Eukaryota / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D023282:Nuclease Protection Assays / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012623:Seawater; D012680:Sensitivity and Specificity
D012337:RNA, Ribosomal, 18S