Ten Years of R&D and Full Automation in Molecular Diagnosis

Abstract

A 10-year experience of our automated molecular diagnostic platform that carries out 91 different real-time PCR is described. Progresses and future perspectives in molecular diagnostic microbiology are reviewed: why automation is important; how our platform was implemented; how homemade PCRs were developed; the advantages/disadvantages of homemade PCRs, including the critical aspects of troubleshooting and the need to further reduce the turnaround time for specific samples, at least for defined clinical settings such as emergencies. The future of molecular diagnosis depends on automation, and in a novel perspective, it is time now to fully acknowledge the true contribution of molecular diagnostic and to reconsider the indication for PCR, by also using these tests as first-line assays.

Evaluation of a quantitative RT-PCR assay for the detection of the emerging coronavirus SARS-CoV-2 using a high throughput system

“…Automated solutions for molecular diagnostics can help handle large numbers of samples and can be scaled to keep pace with fluctuating demand [10][11][12].…”

The epidemiology, diagnosis and treatment of COVID-19

“…The RT-PCR results usually become positive after several days (2-8 days) [27] . Automated solutions for molecular diagnostics can handle large numbers of samples and can be scaled to keep pace with fluctuating demand [28][29][30] . The good analytical performance of a molecular assay for the detection of SARS-CoV-2 on a highthroughput platform, the cobas 6800, was observed with minimal hands-on time, while offering fast and reliable results [31 , 32] .…”

Visceral leishmaniasis in a lung transplant recipient: usefulness of highly sensitive real‐time polymerase chain reaction for preemptive diagnosis

“…After nucleic acids extraction, the amplification is achieved using the Fast Advanced Master Mix (Thermofisher, Waltham, Massachusetts, USA) on the ABI Prism 7900 FAST instrument (Applied Biosystems, Foster City, California, USA) with the following cycling conditions: 2 min at 50°C, 10 min at 95°C, followed by 45 cycles of 1 sec at 95°C and 20 sec at 60°C. The detection limit of this PCR is approximately 10 3 –10 4 DNA copies/mL . Using this PCR, we could detect 8.9 × 10 7 Leishmania kinetoplastic DNA copies/mL in the PB sample of June 23, 2015, which corresponded to 8.9 × 10 3 parasites/mL, as the PCR targets the kinetoplastic DNA present in about 10,000 copies per parasite (Fig.…”

“…The main graphic represents Leishmania detection using a specific real‐time PCR targeting a sequence of the kinetoplastic DNA . The detection limit of the PCR in our molecular diagnostic platform is between 10 3 and 10 4 DNA copies/mL . On June 22, 2015, a thin blood smear stained with Giemsa revealed several leukocytes infected with Leishmania species amastigotes.…”