碩士 / 國立臺灣大學 / 海洋研究所 / 103 / Billfish is an important fishery resource all over the world, and is usually marketed as sashimi, filleted, fish floss and sauce. Most billfish products in Taiwan were only labeled with “billfish” in their ingredients. Therefore, species identification of the billfish products is necessary and will benefit the conservation and regulation of these fishery resourcs. In this study, we developed a real-time PCR approach to identify the six billfish species：Swordfish (Xiphias gladius), Sailfish (Istiophorus platypterus), Black marlin (Istiompax indica), Striped marlin (Kajikia audax), Blue marlin (Makaira nigricans) and Shortbill spearfish (Tetrapturus angustirostris). The whole system contained 14 primers, including 12 species-specific primers and two universal primers designed for discriminating the six billfish from non-billfish species. The 12 species-specific primers showed clear discrimination among the six billfish species, with ΔCt values of the target species ranging between -2.23 ~ 3.93 and the limit of detection is 0.4 ng/μL. In order to minimize the cost and time of whole identification system, we also tested the applicability of the multiplex qPCR by combining several species-specific primers in a single reaction to identify the species. The result showed that the six species can be identified according to the melting curve pattern and Tm values for X. gladius： Tm1 = 75.0 ± 0.0 °C, Tm2 = 83.5 ± 0.0 °C, I. platypterus：Tm1 = 75.2 ± 0.3 °C, Tm2 = 80.5 ± 0.2 °C, I. indica：Tm1 = 77.5 ± 0.0 °C, Tm2 = 80.5 ± 0.0 °C, K. audax：Tm = 80.5 ± 0.0 °C, M. nigricans：Tm1 = 76.0 ± 0.0 °C, Tm2 = 80.5 ± 0.0 °C, T. angustirostris：Tm = 77.0 ± 0.0 °C. An identification protocol for the six billfish species was established in this study. The methods is efficient, sensitive, and cheaper than other conventional methods, which will be helpful for fishery management and protecting the consumer’s rights.
| Identifer | oai:union.ndltd.org:TW/103NTU05279018 |
| Date | January 2015 |
| Creators | Hung-An Chang, 張宏安 |
| Contributors | 蕭仁傑 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 75 |
Page generated in 0.0085 seconds