Detection of two diatoms using sandwich hybridization integrated with nuclease protection assay (NPA-SH)
Tóm tắt
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 × 104 to 1.2 × 106 cells with a detection limit of 42 cells ml−1. The linear range for S. costatum was 6.0 × 104 to 1.0 × 106 cells with a detection limit of 60 cells ml−1. The NPA-SH in this study provides a convenient tool for rapid assessment of HAB species in marine environments.
Tài liệu tham khảo
Cai, Q. S., R. X. Li, Y. Zhen, T. Z. Mi & Z. G. Yu, 2006. Detection of two Prorocentrum species using sandwich hybridization integrated with nuclease protection assay. Harmful Algae 5(3): 300–309.
Frey, A., B. Meckelein, D. Externest & M. A. Schmidt, 2000. A stable and highly sensitive 3,3′,5,5′-tetramethylbenzidine-based substrate reagent for enzyme-linked immunosorbent assays. Journal of Immunology Methods 233: 47–56.
Guillard, R. R. L., 1973. Division rates. In Stein J. R. (ed.), Handbook of Phycological Methods, Vol. 1. Cambridge University Press, Cambridge, 289–312.
Guillard, R. R. L. & J. H. Ryther, 1962. Studies of marine Planktonic diatoms. I. Cyclotellanana Hustedt and Detonula confervacea (Cleve). Canadian Journal of Microbiology 8: 229–239.
Han, X. T., J. Z. Zou & Y. S. Zhang, 2004. Harmful algae bloom species in Jiaozhou Bay and the features of distribution. Marine Sciences 28: 49–54 (in Chinese, with English abstract).
Hasle, G. R., 1978. Settling: The inverted-microscope method. In Sournia, A. (ed.), Phytoplankton Manual. Monographs on Oceanographic Methodology 6. UNESCO, Paris, 88–96.
Hasle, G. R., 1978. Estimating cell numbers: Using the inverted microscope. In Sournia, A. (ed.), Phytoplankton Manual. Monographs on Oceanographic Methodology 6. UNESCO, Paris, 191–196.
Huo, W. Y., Z. M. Yu, J. Z. Zou, X. X. Song & J. H. Hao, 2001. Outbreak of Skeletonema costatum red tide and its relationship with environmental factors in Jiaozhou Bay. Oceanologia et Limnologia Sinica 32: 80–88 (in Chinese, with English abstract).
Jeffrey, S. W. & N. A. Welschmeyer, 1997. Spectrophotometric and fluorometric equations in common use in oceanography. In Jeffrey, S. W., R. F. C. Mantoura & S. W. Wright (eds), Phytoplankton Pigments in Oceanography: Guidelines to Modern Methods. UNESCO, Paris, 661–682.
Jonker, R. R., J. T. Meulemans, G. B. J. Dubelaar, M. F. Wilkins & J. Ringelberg, 1995. Flow cytometry: a powerful tool in analysis of biomass distributions in phytoplankton. Water Science and Technology 32: 177–182.
Koehne, B., G. Elli, R. C. Jennings, C. Wilhelm & H. –W. Trissl, 1999. Spectroscopic and molecular characterization of a long-wavelength absorbing antenna of Ostreobium sp. Biochimica et Biophysica Acta / Bioenergetics 1412: 94–107.
Liu, D. Y., J. Sun, H. T. Chen & L. Y. Zhang, 2003. The phytoplankton community in summer 2001 in Jiaozhou Bay, China. Journal of Ocean University of Qingdao 33: 366–374 (in Chinese, with English abstract).
Manhart, J. R., G. A. Fryxell, C. Villac & L. Y. Segura, 1995. Pseudo-nitzschia pungens and P. multiseries (Bacillariophyceae): nuclear ribosomal DNAs and species differences. Journal of Phycology 31: 421–427.
Okaichi, T., 1989. Red tide problems in the Seto Inland Sea, Japan. In Okaichi, T., D. M. Anderson & T. Nemoto (eds), Red Tides, Biology, Environmental Science and Toxicology. Elsevier, New York, Amsterdam, London, 137–142.
Scholin, C. A., K. R. Buck, T. Britschgi, G. Cangelosi & F. P. Chavez, 1996. Identification of Pseudo-nitzschia australis (Bacillariophyceae) using rRNA-targeted probes in whole cell and sandwich hybridization formats. Phycologia 35: 190–197.
Scholin, C. A., III R. Marin, P. E. Miller, G. J. Doucette, C. L. Powell, P. Haydock, J. Howard & J. Ray, 1999. DNA probes and a receptor-binding assay for detection of Pseudo-nitzschia (Bacillariophyceae) species and domoic acid activity in cultured and natural samples. Journal of Phycology 35: 1356–1367.
Shen, Z. L., 2002. Long-term changes in nutrient structure and its influences on the ecology and environment in Jiaozhou Bay. Oceanologia et Limnologia Sinica 33: 322–331 (in Chinese, with English abstract).
Sieracki, C. K., M. E. Sieracki & J. Yentsch, 1998. An imaging-in-flow system for automated analysis of marine microplankton. Marine Ecology Progress Series 168: 285–296.
Simon, N., L. Campbell, E. Ornolfsdottir, R. Groben, L. Guillou, M. Lange & L. K. Medlin, 2000. Oligonucleotide probes for the identification of three algal groups by dot blot and fluorescent whole-cell hybridization. Journal of Eukaryotic Microbiology 47: 76–84.
Tyrrell, J. V., P. R. Bergquist, P. L. Bergquist & C. A. Scholin, 2001. Detection and enumeration of Heterosigma akashiwo and Fibrocapsa japonica (Raphidophyceae) using rRNA-targeted oligonucleotide probes. Phycologia 40: 457–467.
Venugopalan, C., & H. C. Kapoor, 1997. Single step isolation of plant RNA. Phytochemistry 46: 1303–1305.
Wu, Y. L., & C. X. Zhou, 2003. The research on ecology of HAB in Chinese primary estuaries and bays: Jiaozhou Bay. In Qi, Y. Z., J. Z. Zou & S. Liang (eds), Harmful Algae Blooms in Chinese Coastal Waters. Science Press, Beijing, 136–146 (in Chinese).