This technique allows the detection of the virus in naso- and oropharyngeal samples within 30C40 min. the fight against other infectious diseases. Phage display libraries are a amazingly useful tool that allows the recognition of the best ligands for a given target [75], permitting the building of large libraries consisting of several antibody genes [76]. This Piragliatin type of libraries has been used since 1992 to identify specific monoclonal antibodies (mAb) against particular bacteria or viruses [77,78], and a high percentage of human being therapeutic antibodies have been developed by this technique [79]. In the past, there have been several examples of phage display libraries Piragliatin expressing viral peptides that have successfully inhibited infections, for instance, the ones caused by adenovirus type 2 [80], hepatitis B computer virus [81], hantavirus, sin nombre computer virus [82], and Andes computer virus [83]. This justifies the use of these libraries like a diagnostic and treatment tools of SARS-CoV-2 (Number 1). Open in a separate window Number 1 Uses of phage-display libraries in the diagnostic (1, 2 y 3) and treatment (4, 5, and 6) of SARS-CoV-2. scFv: single-chain variable fragment. S2: spike subunit 2. mAbs: monoclonal antibodies. MERS: Middle-East respiratory syndrome. RBD: receptor-binding protein. 3.1. Bacteriophages mainly because Diagnostic Tools: Phage-Display Libraries Phage display is a powerful technique for the recognition and isolation of peptides or proteins [76]. This technique consists of expressing foreign peptides on the surface of bacteriophages, regularly filamentous bacteriophages isolated from [107] and [108], among others. 4.1. CRISPR-Cas like a Molecular Tool of Diagnostic of COVID-19 In the last few months, several projects related to CRISPR have appeared or have been altered in response to the current crisis caused by the COVID-19 pandemic [109]. All these techniques Piragliatin use primarily the Cas13 and Cas12 proteins because of their capacity to slice single-strands of either DNA or RNA [110]. Most of CRISPR centered techniques have been developed to use Light or RT_Light (Reverse transcription loop-mediated isothermal amplification). This technique was developed to simplify the PCR process, with shorter reaction times and no need for specific products [111]. Besides, these methods can be developed without high technology or troubles, permitting the professionals to perform the diagnostic of the disease directly in the sample collection points. Among this research, we spotlight six Piragliatin main diagnostic checks using CRISPR technology (Table 2): (i) SHERLOCK: Specific High-sensitivity Enzymatic Reporter unLOCKing. This technique uses the RNAse activity of the CRISPR-Cas13a protein, which needs only a small specific RNA guideline [112]. The system was adapted to a simple test against SARS-CoV-2, called STOPCovid (SHERLOCK Screening in One Pot), which counts today with two versions: STOPCovid.v1 and STOPCovid.v2 [113]. Both of them use LAMP technique for RNA amplification and may detect up to 100 viral genome copies per reaction in 45C60 min. STOPCovid.v2 uses magnetic beads to simplify the RNA extraction and reduce its duration [113]. Experts have developed a simple test format that can be performed without complex instrumentation and may detect the computer virus in saliva samples [114]. This method has been clinically validated by a different study group, who have decreased the limit of detection, therefore increasing its level of sensitivity [115].(ii) DETECTR: DNA Endonuclease TargEted CRISPR Trans Reporter. This system uses the CRISPR-Cas12a protein to detect SARS-CoV-2 through its nucleoprotein and envelope genes, based on the method of RT-LAMP, which includes a simultaneous retrotranscription process. This technique allows the detection of the computer virus in naso- and oropharyngeal samples within 30C40 min. The limit of detection is definitely 10 copies per microliter [116].(iii) CARMEN: Combinatorial Arrayed Reactions for Multiplexed Evaluation of Nucleic-acids. This method combines SHERLOCK with microfluidic technology, enabling the analysis of numerous types of samples from patients. The system was developed to detect 169 human-associated viruses, including Cdh15 SARS-CoV-2. Moreover, it can be used for.
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