Nucleic acid (NA) extraction is a basic step for genetic analysis, from scientific research to diagnostic and forensic applications. It aims at preparing samples for its application with biomolecular technologies such as isothermal and non-isothermal amplification, hybridization, electrophoresis, Sanger sequencing and next-generation sequencing. Multiple steps are involved in NA collection from raw samples, including cell separation from the rest of the specimen, cell lysis, NA isolation and release. Typically, this process needs molecular biology facilities, specialized instrumentation and labor-intensive operations. Microfluidic devices have been developed to analyze NA samples with high efficacy and sensitivity. In this context, the integration within the chip of the sample preparation phase is crucial to leverage the promise of portable, fast, user-friendly and economic point-of-care solutions. This review presents an overview of existing lab-on-a-chip (LOC) solutions designed to provide automated NA extraction from human raw biological fluids, such as whole blood, excreta (urine and feces), saliva. It mainly focuses on LOC implementation aspects, aiming to describe a detailed panorama of strategies implemented for different human raw sample preparations.
The effectiveness of a simple PCR protocol performed on paraffin-embedded tissues, obtained from histopathologically and culturally diagnosed cases of dermatophytic pseudomycetoma DPM was tested. The specimens were investigated using previously described primers (DH1L and DH1R) targeting the 18S rDNA gene and amplifying a 183-bp fragment. Microsporum canis was identified from all samples. The PCR protocol described in the present work demonstrated a 100% concordant result comparing the molecular characterisation with phenotypic characterisation of dermatophytes. Molecular biology could represent a valid identification tool in dermatophytic deep infections, when diagnosis cannot be achieved by cultural methods.
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