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Barr: When and how did you all begin showing the RNA extraction free method could match or exceed the sensitivity of standard test?

Hufnagel: Well, we We knew we had the ability to do this in collaboration with the Department of Laboratory Medicine at the Clinical Center. Dr. Karen Frank is the Director of all of the clinical testing that happens at the NIH Clinical Center in Building 10. We were able to synthesize or make synthetic samples in the lab. Bin was able to compare his method to the method used by the CDC [U.S. Centers for Disease Control and Prevention] to show that they acted similarly in a synthetic or an artificial situation where we knew exactly how much virus was going into all the samples. Then we were able to work with the Clinical Center where they were doing the live testing, and they were doing it at the standards that any clinical lab would be doing this as. We were able to work with them to test Dr. Guan’s method with Dr. Frank's laboratory performing the CDC method, side by side, all on the same machines and by the same personnel. We were able to get the samples from the National Institute of Dental and Craniofacial Research Institute (NIDCR) from Dr. Blake Warner. He was one of the doctors in the car line collecting samples from volunteers who were coming in every day. It was a really fantastic collaboration between all these different Institutes and investigators and laboratories so that we could do this. That was really the only way we could have really compared these in what we would call “live testing.”

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Barr: That’s interesting. Were you guys surprised at how well your research went? Were you surprised by your findings at all?  

Hufnagel: Well, you You always set out with a hypothesis, but of course, a lot of experiments simply don't work. We do try a lot of experiments that don't work, and this is one of the ones that did. We're always ecstatic when something sticks. But Bin had a really, really great rationale. Bin actually has a great story he should tell, about his experience with Chelex when he was an instructor for college students.

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Barr: How did you guys get into this research, considering some of the other research you do?

Hufnagel: Well, we We specialize in the extraction of DNA and RNA from patient samples. We extract DNA from patient blood, from saliva, and we have even extracted DNA from pathology samples that have been archived at the National Eye Institute. We have a lot of expertise in the detection of these nucleotides that are basically salts. We understand a lot of the things about handling these so that they don't degrade. That was the sticking point in Bin's mind—that we have these discussions about RNA extraction. We know we have all these kits in the laboratory, and we're always trying to reduce the costs and the workflow. RNA extraction kits are a cost, and they take up space in the lab. We were wondering about some of these direct detection things that might be more broadly applicable to other things. Chelex had been used for DNA previously, and so Bin thought to apply it to RNA, and now we're thinking about how to apply Chelex to other things. Can we detect other kinds of viruses? Can we detect other kinds of RNAs, like our own human RNAs? Can we use Chelex for that sort of benefit? One of the nice things that Bin’s paper also shows is that Chelex permitted for shelf stability when in saliva. It also makes you wonder about being able to collect samples out in the field and then leaving them in your bag and shipping them. Maybe it takes a few days, but you know that the RNA is going to be stable. Whereas in a lot of other circumstances, we try to extract the RNA as quickly as possible so that the enzymes don't get at it and break it down. In this case, Chelex may allow for some extra time to get samples from remote locations into a laboratory, where they can then be safely extracted and detected. This, again, may be for viruses, or other organisms, or even our own DNA and RNA.

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Barr: That's a really exciting possibility.

Hufnagel: YeahYes. We'll be testing out some of these, so we may have to meet with you again soon to talk more about it.

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Barr: I also hope that you all stay safe with your families as well.

Hufnagel: You too.

Guan: Yeah, you You too.

https://pubmed.ncbi.nlm.nih.gov/34396082/

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