Development and validation of a real time PCR-based bioassay for quantification of neutralizing antibodies against human interferon-beta

For many patients suffering from MS, interferon beta (IFNβ) is an effective therapeutic option; however, some patients who receive long-term IFNβ therapy for relapsing-remitting MS (RRMS) develop neutralizing antibodies (NAbs) that affect IFNβ efficacy. It is therefore important to evaluate patients' therapeutic response to IFNβ over time. Myxovirus resistance protein A (MxA), a surrogate marker of individual immunologic response to IFNβ, may be a useful tool for assessing IFNβ immunogenicity. The real-time TaqMan assay for MxA messenger RNA (mRNA) has several distinct advantages, including the ability to amplify and complete quantitative analyses in one step, a high degree of quality control and prior experience and confidence in the field of quantitative viral diagnostics. The real-time TaqMan assay for MxA mRNA can be incorporated as a component of IFNβ therapy to evaluate patients during the course of treatment. Multiple Sclerosis 2007; 13: S49—S52. http://msj.sagepub.com

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Development and validation of TaqMan real-time PCR assays for quantification of chicken adulteration in hamburgers

Journal of Food Composition and Analysis ◽

10.1016/j.jfca.2021.104302 ◽

2021 ◽

pp. 104302

Author(s):

Zahra Sarlak ◽

Saeedeh Shojaee-Aliabadi ◽

Nayebali Rezvani ◽

Hedayat Hosseini ◽

Milad Rouhi ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Pcr Assays ◽

Development And Validation

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Sesame, Pistachio, and Macadamia Nut: Development and Validation of New Allergenic Systems for Fast Real-Time PCR Application

Foods ◽

10.3390/foods9081085 ◽

2020 ◽

Vol 9 (8) ◽

pp. 1085

Author(s):

Martina Torricelli ◽

Elisa Pierboni ◽

Cristina Rondini ◽

Serena Altissimi ◽

Naceur Haouet

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Food Processing ◽

Annealing Temperature ◽

Limit Of Detection ◽

Food Samples ◽

Support Tool ◽

Macadamia Nut ◽

Real Market ◽

Development And Validation

Food allergy is a worldwide health problem that concerns infants to adults. The main health risk for sensitised individuals is due to the presence of traces of allergens as the result of an accidental contamination during food processing. The labelling of allergens such as sesame, pistachio, and macadamia nut on food products is mandatory according to Regulation (EU) N. 1169/2011; therefore, the development of suitable and specific analytical methodologies is advisable. The aim of this study was to perform a multi-allergen real-time PCR system that works well in fast mode at the same annealing temperature and with the same thermal profile. The real-time PCR was developed designing new, specific, and efficient primer and probe systems for the 2S albumingene for sesame and pistachio and for the vicilin precursorgene for macadamia nut. These systems were subjected to a robust intra-laboratory qualitative validation process prior to their application, by DNA extraction and fast real-time PCR, on some real market samples to reproduce a potential allergen contamination along the food chain. The developed system results were specific and robust, with a sensible limit of detection (0.005% for sesame; 0.004% for pistachio; 0.006% for macadamia nut). The performance and the reliability of the target systems were confirmed on commercial food samples. This molecular approach could be used as a screening or as a support tool, in association with the other widespread monitoring techniques (such as ELISA).

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