Automatic system for high-throughput and high-sensitivity diagnosis of SARS-CoV-2

Abstract

The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has had severe consequences for health and the global economy. To control the transmission, there is an urgent demand for early diagnosis and treatment in the general population. In the present study, an automatic system for SARS-CoV-2 diagnosis is designed and built to deliver high specification, high sensitivity, and high throughput with minimal workforce involvement. The system, set up with cross-priming amplification (CPA) rather than conventional reverse transcription-polymerase chain reaction (RT-PCR), was evaluated using more than 1000 real-world samples for direct comparison. This fully automated robotic system performed SARS‐CoV‐2 nucleic acid-based diagnosis with 192 samples in under 180 min at 100 copies per reaction in a “specimen in data out” manner. This throughput translates to a daily screening capacity of 800–1000 in an assembly-line manner with limited workforce involvement. The sensitivity of this device could be further improved using a CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-based assay, which opens the door to mixed samples, potentially include SARS-CoV-2 variants screening in extensively scaled testing for fighting COVID-19. Supplementary Information The online version contains supplementary material available at 10.1007/s00449-021-02674-9.

Comparación de dos métodos isotérmicos para la detección de SARS-COV-2 en Callao, Perú

“…The evaluated results show that both techniques have adequate and equivalent values of sensitivity (RT-LAMP: 82.8%, CPA: 83%) and specificity (RT-LAMP and CPA: 91.5%), as well as a high concordance (88%) between both techniques. Our results are in line with previous studies such as that of Lu et al [5] showed that the CPA technique shows high sensitivity and specificity versus the gold standard, been able to detect from 100 copies of ARN per reaction tube. On the other hand, Rai et al [16] based on a systematic review of different molecular techniques for SARS-CoV-2 detection found that RT-LAMP is capable to detect from 4 copies/uL.…”

“…The importance of applying molecular screens is that they can detect the presence of virus or its remnants [5] at any stage of the disease, because these screens have been shown to detect high, medium and low viral loads, unlike serological tests because performance is influenced by symptomatology, days of infection, or virus lineage and they are not confirmatory tests for the disease [13][14][15] .…”

“…In view of this problem, there is an urgent need to use alternative cost-effective techniques to aid in the diagnosis of COVID-19, such as the reverse transcription loop-mediated isothermal amplification (RT-LAMP) [3,4] and cross-priming amplification (CPA) methods [5] . Currently, both methods are used in Callao for the mass screening of SARS-CoV-2; nonetheless, it is uncertain if they are equivalent techniques, possibly leading to misguided results.…”

Multiplex detection of bacterial pathogens by PCR/SERS assay

“…22,[30][31][32] In the realm of molecular diagnostics, a sensitivity of 100 copies is considered quite good for many applications. [33][34][35] For highly sensitive diagnostic tests such as the detection of trace amounts of pathogens, even greater sensitivity might be necessary to detect lower quantities of targets, such as singledigit or even single-copy detection. 36 To improve the sensitivity of the PCR/SERS assay for DNA targets, the following two factors should be considered: (i) optimization of the PCR amplification, including the primer design, PCR thermal cycle conditions, PCR buffer and components; and (ii) optimization of the SERS-active substrates that could enhance the Raman signal intensities.…”

A high-throughput fully automatic biosensing platform for efficient COVID-19 detection

“…The system involves 6 main steps, and 6 different equipment are needed to complete each step. Another automatic system having better compactness (one single floor-type equipment) and higher throughput has been proposed that can carry out SARS-CoV-2 detection of 192 samples under 180 min at detection limit of 100 copies per reaction ( Lu et al, 2022 ). In this work, we propose a desktop type automatic detection system that can detect SARS-CoV-2 of 384 samples in around 30 min with detection limit of 100 TCID 50 /ml.…”