Infectious Diseases Detection Platform: COVID-19

Friday, May 15, 2020

The first known case of novel (new) Coronavirus occurred on December 1st, 2019. On December 31, 2019, Chinese authorities notified the World Health Organization of multiple flu-like cases in Wuhan City, Hubei Province, China and which continues to expand is being closely monitored. On February 11, 2020, the World Health Organization named the disease Coronavirus disease 2019 (abbreviated “COVID-19”). Chinese health officials have reported over 80,000 cases of COVID-19 in China, with the virus reportedly spreading rapidly from person-to-person in parts of that country. The United States reported the first confirmed instance of person-to-person with this virus on January 30, 2020, and so far has exceeded 100,000 confirmed cases. The U.S. government is working closely with public health partners, to respond to this public health threat. The public health’s goals are to detect quickly and minimize the spread and impact of this virus in the United States and globally.

Recently, most of the standard viral detection methods around the world are using real-time PCR. However, the operation and interpretation of real-time PCR test results require professional or well-trained technicians and false-positive results are often occurring.

BiOptic, Inc. a Taiwan based R&D company with a branch office in La Canada Flintridge, California has developed a total solution, which includes Direct "RT-PCR+PCR" reagents,Qamp-mini PCR thermal cycler (for 8-samples) with a pre-programmed chip, and the portable Capillary Gel Electrophoresis instrument (Qsep1-Lite) to solve the problems associated with traditional real-time PCR systems (Figure 1).

Figure 1. Quick and Easy Steps to Get Results at High Sensitivity Confirming the Real-time PCR Results

Our detection platform utilizes in house developed Direct "RT-PCR+PCR" reagents, which eliminates the need for nucleic acid extraction, where it provides a unique solution for rapid pathogen detection with reproducible results by nontechnical operators. Also, it resolves the false positive issues associated with real-time PCR systems, since our platform has much higher sensitivity and provides better detection capabilities. Furthermore, BiOptic’s detection platform should be suitable as a reconfirm platform after real-time PCR (Figure 2) increasing the reliability of the diagnosis of the disease more efficiently.

Figure 2. Higher Detection Sensitivity results produced by Qsep1-Lite capillary gel electrophoresis system

BiOptic’s Detection Platform has been used through our collaborator in China for the COVID-19 testing application. With limited samples, the results showed the BiOptic test is more sensitive in picking up true positive sample than other RT-PCR testing methods (Figure 2). Furthermore, our test is much simpler to perform.

BiOptic Detection Platform includes mixing the swab with our inhouse developed lysis buffer and transfer of 2 microliters of the lysed sample directly onto our portable device performing the tests automatically and providing test results within an hour. BiOptic’s compact detection platform can provide fast, accurate, and cost-effective results that should be suitable for decentralized testing application of COVID-19.

BiOptic Detection Platform provides:

One-Step RT PCR amplification. There is no need for RNA extractions and purifications. Buccal or nasal swabs are immersed into a lysis buffer that liberates the nucleic acids. The RNA in these lysates can then be used to set up the Reverse Transcriptase PCR Amplifications.
Reverse Transcriptase PCR Amplifications are run on the Qamp-mini PCR System. There is no need for costly Real-Time PCR instruments based on the use of fluorescent Taqman probes (or equivalent), that read and record increases in fluorescence during the 35+ PCR cycles.
PCR products that have been generated after the full number of cycles by the Qamp-mini are then analyzed by Capillary Gel Electrophoresis (CGE) utilizing the fully automated Qsep1-Lite Analysis System. Results for up to 8 samples can be obtained quickly.

In a couple of cases, Real-Time RT-PCR assays had been run and had shown questionable results. Analysis of the same sample using BiOptic’s technology demonstrated that COVID-19 RNA sequences were present in these samples as shown in the CGE tracings below (Figure 2). These results indicate that this technology can be more sensitive than standard RT-PCR assays.

(A) Covid-19 detection kit (Company A): Red line: Positive control, Blue line: Negative control, Green line: Reconfirm Negative Sample No.1 and Yellow line: Reconfirm Negative Sample No.2. Red arrow indicates the amplicon signals detected by Qsep Series.

(B) Covid-19 detection kit (Company B): Red line: Positive control, Blue line: Negative control, Green line: Reconfirm Negative Sample No.1

The above analysis demonstrates the sensitivity of our detection platform. We used Qsep Series (Qsep1-Lite) to analyze samples that were determined as negative results by qPCR. It is obvious that the signal peak can be seen in the corresponding target area. This advantage can effectively reduce the issue of qPCR false negatives and each analysis takes only 3 minutes.

Everything that one needs for COVID-19 testing are provided in a portable kit that can be taken into the field. You are not confined to having to run the assays in a fully equipped laboratory. Lysis buffers, PCR primers and reagents, Qamp-mini PCR Thermal Cycler, and the Qsep1-Lite CGE Analysis system are all included in a case that can be loaded into the trunk of a car or an SUV (Figure 3).