Identification of Ingredient in Mullet Roe Products by the Real-Time PCR Method
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Identification of Ingredient in Mullet Roe Products by the Real-Time PCR Method Je-Hung Kuo 1 & Hsiu-Wei Tsuei 1 & Zih-Ling Jia 1 & Che-Yang Lin 1 & Yuan-Hsin Chang 1 & Bo-Lin Chen 1 & Jung Kuan 1 & Hsu-Yang Lin 1 & Lih-Ching Chiueh 1 & Daniel Yang-Chih Shih 1 & Hwei-Fang Cheng 1
Received: 15 May 2016 / Accepted: 19 September 2017 / Published online: 27 October 2017 # Springer Science+Business Media, LLC 2017
Abstract Mullet roe is a product of high economic value in a number of Asian countries, particularly Taiwan. However, actual mullet roe is commonly adulterated by the addition of other species, such as escolar and oilfish. The purpose of this study was to develop a method to detect the ingredient of mullet roe products. Based on the TaqMan real-time PCR assay, we designed the specific primers-probe set (mullet) that targets the mitochondrial 16S ribosomal RNA gene. Meanwhile, the positive amplification control is designed based on the eukaryotic 18S rRNA gene. The PCR amplicon used to identify fish species in processed roe products is smaller than 200 bp. Method specificity was evaluated by analyzing tissue samples of 29 food fish and 9 puffer fish species. No indications of cross-reactivity toward non-target species were observed. Sensitivity and linearity tests were conducted using five-fold serial dilutions of target DNA from processed mullet roe, and we determined that our proposed method has a sensitivity of 1.2 ng. Further tests on a random survey of commercial fish roe products demonstrated the efficacy of the technique in the detection of mullet DNA. The real-time PCR methods developed in this study could be used to verify the labeling of actual mullet roe products.
Keywords 16S rRNA gene . Real-time PCR . Mullet . Food authentication
* Hsu-Yang Lin [email protected]
1
Division of Research and Analysis, Taiwan Food and Drug Administration, Taipei, Taiwan (R.O.C.)
Introduction Mugil cephalus is a euryhaline marine fish that tolerates a wide range of salinities, from freshwater to hypersaline conditions, and can persist at water temperatures of 12–25 °C (Thomson 1966; Kuo et al. 1973; Collins 1985; VegaCendejas and Hernández de Santillana 2004). M. cephalus is distributed globally in coastal and estuarine waters of subtropical and tropical zones (between latitudes 42°N and 42°S) (Thomson 1966). With the coming of the northeast monsoon, M. cephalus growing in the rivers of mainland China enter the sea and swim to spawning grounds in Southeast Asia to mate. Around the time of the winter solstice (21 or 22 Dec.), M. cephalus makes an annual migration from the coastal waters of northern China to the waters southwest of Taiwan (Chen and Su 1986; Hunag and Su 1989). Ten days before and after the winter solstice is the best period to catch M. cephalus (Tung 1981; Hunag and Su 1989). M. cephalus is known in Taiwan as Bblack gold^. The most popular product processed from this fish is M. cephalus roe (Local name, BWuyuzi^), due to its nutritional value (Liao 1981; Barra et al. 2008; Scano et al. 2008, 2009;
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