Comparison of <scp>DNA</scp> extraction methods for entomological origin identification of honey using simple additive weighting method

Meng

et al. 2020

CyTA - Journal of Food

The objective of this study was to establish a real-time LAMP assay for authentication of rape (Brassica napus) honey to protect consumers from commercial honey adulteration. The LAMP primers targeting the internal transcribed spacer (ITS) of Brassica napus were designed, and its specificity was tested. The LAMP reaction temperature was also optimized, and the detection limit of the LAMP assay was determined with a serial dilution of genomic DNA from the seeds of Brassica napus. The results showed that the real-time LAMP assay can accurately and specifically detect the rape component in honey, and the detection limit was 10 pg genomic DNA of Brassica napus. Data on monofloral honey samples indicate that the real-time LAMP assay was 100% in concordance with the reported TaqMan TM PCR assay. This study provides a promising solution for facilitating the authentication of rape honey in food retail market.Desarrollo de un ensayo LAMP en tiempo real para la autenticación de la miel monofloral utilizando miel de colza RESUMEN El presente estudio se propuso establecer un ensayo LAMP en tiempo real destinado a autenticar la miel de colza (Brassica napus) para proteger a los consumidores de la adulteración comercial de la miel. Con esta finalidad se diseñaron cebadores LAMP dirigidos al espaciador transcrito interno (ITS) de Brassica napus y se probó su especificidad. Además, se optimizó la temperatura de reacción del LAMP, determinándose el límite de detección del ensayo LAMP con una dilución en serie de ADN genómico de las semillas de Brassica napus. Los resultados mostraron que dicho ensayo en tiempo real puede detectar con precisión y especificidad el componente de colza en la miel; el límite de detección fue de 10 pg de ADN genómico de Brassica napus. Los datos obtenidos de las muestras de miel monofloral dan cuenta de que el ensayo LAMP en tiempo real concordaba al 100% con el ensayo de PCR TaqMan TM notificado. Este estudio ofrece una solución prometedora para facilitar la autenticación de la miel de colza en el mercado minorista de alimentos.

Section: Discussionmentioning

confidence: 99%

“…Genomic DNA from the honey samples was extracted by the CTAB-based method modified based on previous reports (Kek et al, 2018). Briefly, 10 mL of honey sample and 35 mL of deionized water were mixed in a 50-mL tube and incubated at 65°C to reach full dissolution, and then centrifuged at 2656 × g for 30 min.…”

Section: Genomic Dna Extractionmentioning

confidence: 99%

Development of a real-time LAMP assay for monofloral honey authentication using rape honey

Meng

et al. 2020

CyTA - Journal of Food

Honey Analysis - New Advances and Challenges

“…Waiblinger et al [177] described an in-house and interlaboratory validation of a DNA extraction method from pollen in a unifloral rape honey with several multifloral honeys. While most DNA studies on honey have focused on botanical species identification, Kek et al [54,55] have determined the best DNA extraction of bees in honey and introduced entomological identification of honey based on bee mitochondrial 16S rRNA and COI gene sequences. Table 8 presents other methods which included instrumental and improvised analytical procedures used for honey identification and authentication.…”

Section: Molecular Techniquesmentioning

confidence: 99%

A Review on Analytical Methods for Honey Classification, Identification and Authentication

Chin¹,

Sowndhararajan²

2020

Authentication of food products is of great concern in the context of food safety and quality. In recent years, interest in honey authenticity in relation with botanical or geographical origin and adulteration has increased. Honey is a ready-to-eat natural food with high nutritional content and gives many health benefits. Authentication of honey has primary importance for both industries and consumers in combatting common honey frauds in the form of mislabeling of honey origin and adulteration with sugar or syrups. Various analytical methods are used for detecting original honey. With a diverse range of equipment and techniques, the conventional analytical methods are still being used in association with advanced techniques as they are part of preliminary screening, processing and product standards. Most of the analytical methods provide indications of pollen distribution, physicochemical parameters and profile analysis of phenolic, flavonoid, carbohydrate, amino acids, aroma and individual marker components. This review provides an overview and summary of instrumental and analytical methods available for honey authentication from conventional to recent molecular techniques. It is useful as a guide to choosing appropriate method for analysis, classification and authentication of honey.

“…An efficient and reliable DNA extraction method must be effective in yielding adequate amount of high-quality extracted DNA and suitable for subsequent downstream molecular analyses such as conventional/end point PCR, real-time PCR, and DNA microarrays [ 16 , 17 , 18 ]. Various studies that have evaluated and compared DNA extraction methods on different subject matters are available [ 19 , 20 , 21 , 22 ].…”

Section: Introductionmentioning

confidence: 99%

Optimum DNA Extraction Methods for Edible Bird’s Nest Identification Using Simple Additive Weighting Technique

Quek

Chin

Tan

2021

Foods

Self Cite

A simple additive weighting (SAW) technique was used to determine and compare the overall performance of five DNA extraction methods from conventional (SDS method) to commercial kits (Qiagen, Wizard, and NucleoSpin) for identifying origins of edible bird’s nest (EBN) using end-point polymerase chain reaction (PCR). A hybrid method (SDS/Qiagen) which has been developed by combining the conventional SDS method with commercialised Qiagen was determined as the most suitable in terms of speed and cost-effectiveness. The determination of optimum extraction method was by the performances on efficiency and feasibility, extracted DNA concentration, purity, PCR amplifiability, handling time and safety of reagents used. The hybrid SDS/Qiagen method is less costly compared to the commercial kits and offered a more rapid alternative to the conventional SDS method with significant improvement in the yield, purity and PCR amplifiability. The developed hybrid SDS/Qiagen method provides a more practical alternative over the lengthy process using conventional method and expensive process using commercial kits. Using the simple additive weighting (SAW) technique and analysis, the Qiagen method is considered the most efficient and feasible method without consideration of cost as it yielded the purest extracted DNA and achieved the highest PCR amplifiability with the shortest turnaround time.