Introduction

A diagnostic kit for the multiple detection of 4 viruses of the Coronaviridae family: HCoV, SARS-CoV, MERS-CoV and the viral strain SARS-CoV-2 which caused a surge of the disease known as COVID-19. From a more general point of view, diagnostic methods and devices intended for the detection of viruses of the Coronaviridae family. In fact, in cases of epidemics or pandemics such as those caused by viruses of the Coronaviridae family (also called Coronavirus), such as SARS-CoV-2, SARS-CoV, MERS-CoV, HCoV, it is important to be able to establish the health conditions of potential suspected cases quickly and effectively, not only in order to save lives, but also to have knowledge of the spread of the infection and take the necessary health measures.

Technical features

The kit provides a “One-Step” method of quantitative gene amplification after retro-transcription of the viral genome (rRT-PCR).The idea behind the invention is based with the aid of the rRT-PCR test, for the quantitative detection of the nucleic acid of a plurality of viruses representative of some pathologies related to the Coronaviridae family, in respiratory samples of the upper and lower tract (such as nasopharyngeal or oropharyngeal swabs), sputum, lower respiratory tract aspirate, bronchial alveolar lavage, and nasopharyngeal lavage / aspirate or nasal aspirate, collected from individuals suspected of COVID-19 by their healthcare provider.

Possible Applications

• The current “pandemic” scenario (COVID-19) requires reliable diagnostic tests, such as that of the present invention, in order to attend with the necessary therapeutic decision-making process;
• The rRT-PCR method allows to exclude the presence of contamination or false positives (exclusion of virus fragments in host endothelial cells);
• The rRT-PCR method allows to exclude the presence of false negatives (exclusion of undetectable virus fragments copies).

Advantages

• An enzymatic stabilizer is used: does not need to prepare the mix for PCR;
• The selected primers are targeted to the short fragment, so the time for the PCR phase is less than 1 hour;
• The amplification cycle takes place in a “One-Step” procedure through MPL1 and MPL2;
• In this single process, both false negatives and false positives are highly identified.