In-house Validation of a DNA Extraction Protocol from Honey and Bee Pollen and Analysis in Fast Real-Time PCR of Commercial Honey Samples Using a Knowledge-Based Approach

Torricelli, Martina; Pierboni, Elisa; Tovo, Gloria Raquel; Curcio, Ludovica; Rondini, Cristina

doi:10.1007/s12161-016-0539-x

Cited by 11 publications

(12 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…DNA was isolated from loose Poaceae pollen grains and aerobiological samples on different supports (melinex tape or Petri dish) using a modified CTAB method (Torricelli et al., ). Sterile distilled water was used as negative control for DNA extraction in each session.…”

Section: Methodsmentioning

confidence: 99%

“…After exposure, they were simply plugged and sealed with parafilm until the beginning of the extraction protocol, when the first reagent was added directly in them. DNA was isolated from loose Poaceae pollen grains and aerobiological samples on different supports (melinex tape or Petri dish) using a modified CTAB method (Torricelli et al, 2016). Sterile distilled water was used as negative control for DNA extraction in each session.…”

Section: Preparation Of Aerobiological Samples Dna Extraction Andmentioning

confidence: 99%

“…Pollen samples collected from honeybees, bee nests, and honey have been analyzed with barcoding and metabarcoding approaches in a number of studies, using both nuclear (e.g., ITS2; Keller et al., ; Richardson, Lin, Sponsler, et al., ; Sickel et al., ;. actin ; Torricelli, Pierboni, Tovo, Curcio, & Rondini, ) and plastidial (e.g., rbcL , trnL , matK ) (Bruni et al., ; Galimberti et al., ; Hawkins et al., ; Richardson, Lin, Quijia, et al., ; Valentini, Miquel, & Taberlet, ) marker regions. Pollen DNA extraction and downstream processing are less common with aerobiological samples, which contain a small but very complex amount of biologic material.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

New biomolecular tools for aerobiological monitoring: Identification of major allergenic Poaceae species through fast real‐time PCR

Ghitarrini

Pierboni

Rondini

et al. 2018

Ecology and Evolution

Self Cite

View full text Add to dashboard Cite

Grasses (Poaceae) are very common plants, which are widespread in all environments and urban areas. Despite their economical importance, they can represent a problem to humans due to their abundant production of allergenic pollen. Detailed information about the pollen season for these species is needed in order to plan adequate therapies and to warn allergic people about the risks they take in certain areas at certain moments. Moreover, precise identification of the causative species and their allergens is necessary when the patient is treated with allergen‐specific immunotherapy. The intrafamily morphological similarity of grass pollen grains makes it impossible to distinguish which particular species is present in the atmosphere at a given moment. This study aimed at developing new biomolecular tools to analyze aerobiological samples and identifying major allergenic Poaceae taxa at subfamily or species level, exploiting fast real‐time PCR. Protocols were tested for DNA extraction from pollen sampled with volumetric and gravimetric methods. A fragment of the matK plastidial gene was amplified and sequenced in Poaceae species known to have high allergological impact. Species‐ and subfamily‐specific primer–probe systems were designed and tested in fast real‐time PCRs to evaluate the presence of these taxa in aerobiological pollen samples. Species‐specific systems were obtained for four of five studied species. A primer–probe set was also proposed for the detection of Pooideae (a grass subfamily that includes also major cereal grains) in aerobiological samples, as this subfamily includes species carrying both grass allergens from groups 1 and 5. These, among the 11 groups in which grass pollen allergens are classified, are considered responsible for the most frequent and severe symptoms.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Preparation Of Aerobiological Samples Dna Extraction Andmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

New biomolecular tools for aerobiological monitoring: Identification of major allergenic Poaceae species through fast real‐time PCR

Ghitarrini

Pierboni

Rondini

et al. 2018

Ecology and Evolution

Self Cite

View full text Add to dashboard Cite

show abstract

“…Methods have been described to detect specific floral pollen DNA fragments in honey using PCR and metabarcoding (Jain et al 2013;Lalhmangaihi et al 2014;Guertler et al 2014;Bruni et al 2015;Soares et al 2015;Hawkins et al 2015;Torricelli et al 2016;Prosser and Hebert 2017) and recently advocated as a marker for mānuka honey (MPI 2017c). These markers indicate the presence but not the amount of mānuka honey in a sample.…”

Section: Dnamentioning

confidence: 99%

A Critical Review of the Factors Available for the Identification and Determination of Mānuka Honey

Burns

Dillon

Warren³

et al. 2018

Food Anal. Methods

View full text Add to dashboard Cite

show abstract

“…Finally the pollen was suspended with 0.5ml of CTAB extraction. The performance of pretreatments was validated (Torricelli et. al., 2016).…”

mentioning

confidence: 99%

Molecular Biological Methods in Environmental Engineering

Li¹,

Xia²,

Zhang³

et al. 2017

Water Environment Research

View full text Add to dashboard Cite

Microorganisms can respond fast to the environmental changes, and well suited for environmental assessment. DNA and RNA are generally extracted by extraction kits. After extraction, molecular biological methods including genomics, transcriptomics, proteomics and metabolomics are applied to characterize the genetic and functional diversity of microorganisms. Quantitative PCR, FISH and mass sequence of amplicons are also described. This review provides environmental engineers and microbiologists an overview of important advancements in molecular techniques and highlights the application of these methods in diverse environments.

show abstract

In-house Validation of a DNA Extraction Protocol from Honey and Bee Pollen and Analysis in Fast Real-Time PCR of Commercial Honey Samples Using a Knowledge-Based Approach

Cited by 11 publications

References 17 publications

New biomolecular tools for aerobiological monitoring: Identification of major allergenic Poaceae species through fast real‐time PCR

New biomolecular tools for aerobiological monitoring: Identification of major allergenic Poaceae species through fast real‐time PCR

A Critical Review of the Factors Available for the Identification and Determination of Mānuka Honey

Molecular Biological Methods in Environmental Engineering

Contact Info

Product

Resources

About