Detection of two diatoms using sandwich hybridization integrated with nuclease protection assay (NPA-SH)

• Published in: Hydrobiologia Vol. 575; no. 1; pp. 1 - 11
• Main Authors: Zhen, Yu, Yu, Zhigang, Cai, Qingsong, Mi, Tiezhu, Li, Rongxiu
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 2007; Springer Nature B.V
• Subjects: Animal and plant ecology; Animal, plant and microbial ecology; Bacillariophyceae; Bacteria; Biological and medical sciences; Brackish; Chaetoceros curvisetus; Fundamental and applied biological sciences. Psychology; General aspects; Hybridization; Marine; Marine environment; Skeletonema costatum; Synecology; INW, China, People's Rep., Shandong Prov., Jiaozhou Bay; Skeletonema costatum; Sandwich hybridization integrated with nuclease protection assay (NPA-SH); Enzyme; Algae; Skeletonema costatum; Esterases; Hybridization; Assay; Heterokontophyta; Aquatic environment; Bacillariophyta; Hydrobiology; Hydrolases; Chaetoceros curvisetus; Nuclease; Detection; Protection; Thallophyta
• ISSN: 0018-8158; 1573-5117
• DOI: 10.1007/s10750-006-0278-x

Nuclease protection assay (NPA) probes were designed to target the rRNA of Chaetoceros curvisetus and Skeletonema costatum, and quantitative sandwich hybridization integrated with nuclease protection assay (NPA-SH) was developed to detect C. curvisetus and S. costatum in culture and field samples in Jiaozhou Bay, China. The specificity and validity of the NPA-SH technique were tested with cultured pure strains, mixed strains and field samples, and by comparison with that of microscopy observation. The linear detection range for C. curvisetus was 4.2 × 10^sup 4^ to 1.2 × 10^sup 6^ cells with a detection limit of 42 cells ml^sup -1^. The linear range for S. costatum was 6.0 × 10^sup 4^to 1.0 × 10^sup 6^ cells with a detection limit of 60 cells ml^sup -1^. The NPA-SH in this study provides a convenient tool for rapid assessment of HAB species in marine environments.[PUBLICATION ABSTRACT]