Specific Detection of Potentially Allergenic Kiwifruit in Foods Using Polymerase Chain Reaction

Abstract

Kiwifruit (Actinidia deliciosa and Actinidia chinensis) is allergenic to sensitive patients, and, under Japanese regulations, it is one of the food items that are recommended to be declared on food labeling as much as possible. To develop PCR-based methods for the detection of trace amounts of kiwifruit in foods, two primer pairs targeting the ITS-1 region of the Actinidia spp. were designed using PCR simulation software. On the basis of the known distribution of a major kiwifruit allergen (actinidin) within the Actinidia spp., as well as of reports on clinical and immunological cross-reactivities, one of the primer pairs was designed to detect all Actinidia spp. and the other to detect commercially grown Actinidia spp. (i.e., kiwifruit, Actinidia arguta, and their interspecific hybrids) except for Actinidia polygama. The specificity of the developed methods using the designed primer pairs was verified by performing PCR experiments on 8 Actinidia spp. and 26 other plants including fruits. The methods were considered to be specific enough to yield target-size products only from the target Actinidia spp. and to detect no target-size products from nontarget species. The methods were sensitive enough to detect 5-50 fg of Actinidia spp. DNA spiked in 50 ng of salmon testis DNA used as a carrier (1-10 ppm of kiwifruit DNA) and 1700 ppm (w/w) of fresh kiwifruit puree spiked in a commercial plain yogurt (corresponding to ca. 10 ppm of kiwifruit protein). These methods would be expected to be useful in the detection of hidden kiwifruit and its related species in processed foods.

Physical and compositional characteristics of ‘mitsuko’ and local hardy kiwifruits in Japan

“…Quinic acid (g 100 g -1 FW) L-malic acid (g 100 g -1 FW) Citric acid (g 100 g -1 FW) Actinidin may display allergenic properties (Fiocchi et al, 2004;Taguchi et al, 2007), whereas it is functional for the digestion (Préstamo, 1995;Rush et al, 2002). Recently, actinidin extract, such as Zyactin (digestive aids; Vital Food Processors Ltd, Auckland, New Zealand), has been commercialized.…”

Yield and amplificability of different DNA extraction procedures for traceability in the dairy food chain

“…The polymerase chain reaction (PCR) is considered the most acknowledged and adaptable DNA-based assay technique, capable to reproduce the genetic information even from strongly damaged DNA sequence. This analytical method is considered very efficient to detect adulteration in foods (Feligni et al, 2005), to monitor the presence of contaminants or pathogens (Cremonesi et al, 2007;Khare et al, 2004), allergens (Taguchi et al, 2007), genetically modified material (Lipp et al, 2005;Nemeth et al, 2004) and to reveal species substitution (Abdel-Rahman & Ahmed, 2007;Di Pinto, Conversano, Forte, Novello, & Tantillo, 2004;López-Calleja Díaz, González Alonso, Fajardo, Martin et al, 2007;López-Calleja et al, 2005;Mašková & Paulíčková, 2006). A possible application of PCR to internal quality traceability along the dairy food chain is the detection of the addition of cheaper milk to high quality and more expansive milk (Bania, Ugorski, Polanowski, & Adamczyk, 2001;Calvo, Osta, & Zaragoza, 2002;López-Calleja, González, Fajardo, Rodríguez et al, 2007;Mafra, Roxo, Ferreira, & Oliveira, 2007;Maudet & Taberlet, 2001).…”

A Comprehensive Review on Kiwifruit Allergy: Pathogenesis, Diagnosis, Management, and Potential Modification of Allergens Through Processing

“…Taguchi et al. () developed a PCR method targeting ITS‐1 region in kiwifruit by designing two primer pairs (F123 and R178). The results showed that this method is able to detect amounts ranging from 5 to 50 fg of kiwifruit allergens.…”