Over the last few years, we’ve all been given a valuable lesson in both the promise and limitations of advanced molecular biology methods for clinical diagnostics. Polymerase chain reaction (PCR) was held up as the “gold standard” of COVID-19 testing, but the cost, complexity, and need for advanced instrumentation and operators with specialized training made PCR difficult to scale to the levels demanded by a pandemic.
There are other diagnostic methods, of course, some of which don’t have all the baggage of PCR. RT-LAMP, or reverse transcriptase loop-mediated amplification, is one method with a lot of promise, especially when it can be done on a cheap open-source instrument like qLAMP. For about 50€, qLAMP makes amplification and detection of nucleic acids, like the RNA genome of the SARS-CoV-2 virus, a benchtop operation that can be performed by anyone. LAMP is an isothermal process; it can be done at one single temperature, meaning that no bulky thermal cycler is required. Detection is via the fluorescent dye SYTO 9, which layers into the base pairs inside the amplified DNA strands, using a 470-nm LED for excitation and a photodiode with a filter to detect the emission. Heating is provided by a PCB heater and a 3D-printed aluminum block that holds tubes for eight separate reactions. Everything lives in a 3D-printed case, including the ESP32 which takes care of all the housekeeping and data analysis duties.
With the proper test kits, which cost just a couple of bucks each, qLAMP would be useful for diagnosing a wide range of diseases, and under less-than-ideal conditions. It could also be a boon to biohackers, who could use it for their own citizen science efforts. We saw a LAMP setup at the height of the pandemic that used the Mark 1 eyeball as a detector; this one is far more quantitative.
Yeah, the side effects of stupidity did so much harm to our society that it will take decades for it to heal, if ever.
Robert F. Kennedy Jr.?
> Aaaand cue the antivax nutjobs.
Because criticizing the actions surrounding a single situation is exactly the same as criticizing an entire field of medicine, and crazy as well?
Those words don’t apply, the language doesn’t work like that. You’re spouting nonsense, while making the fundamental attribution error. Look that up, there’s a Wikipedia page on it.
Until your side stops reciting talking points from rote and starts addressing the targeted criticisms, your side will never be taken seriously.
And there it is.
If i’d had the option to vax i wouldn’t have because i’d had the scourge but couldn’t get back to my family without it. I had the antibodies surely.
One really should go beyond NPR and the Daily Mail or the government mouth pieces. Anyone could use the CDC death certificate charts from the beginning and watch the errors in nearly real-time (about 10 day lag). Some of use did. https://www.youtube.com/watch?v=qrIeYpCp0fc
And another one.
Yeah, charts are hard for you. Here. Tell me why schools should have been closed. https://www.cdc.gov/nchs/nvss/vsrr/covid_weekly/
Schools should have been closed because putting together a group of people in a small area creates an ideal condition for a virus to spread. The people in this case are children, who, for a long time, could not be vaccinated, and who then take their infection back home to their families. Next question, please.
Just be careful, these things tend to have an awfully high false positive rate, so remember, if they aren’t presenting symptoms of a disease, don’t count them as a “case”. I’m sure there’s a lot of biohacking and research that this can enable, but the catastrophic global over-reaction to that mildly nasty cough in 2020/21 shows the folly of trusting PCR/LAMP technologies as a standalone method for diagnosis.
I’m generally not a fan of nucleic acid amplification for clinical diagnosis. PCR, LAMP, whatever — it’s like putting the output from copying a blank piece of paper back onto a copier over and over; eventually you’ll amplify random noise and get a “signal”.
That said, LAMP and RT-LAMP have a LOT of potential for non-clinical public health use cases, like monitoring waste water for viruses and bacteria. LAMP in general and low-cost devices like this in particular make finding reservoirs of disease or surveillance of microorganism shed by large populations much easier. Which is why I wrote this up.
Exactly! That’s why real-time analysis is crucial, because if you check the result (yes/no) after 45 minutes, everything might appear positive. However, with real-time measurements, you can start characterizing after what duration your risk of “noise amplification” becomes significant, and discriminating by the amplification curve of each sample.
Another challenge with LAMP (which can also be seen as an advantage) is that it creates massive amounts of DNA quite rapidly, far more than PCR, and this DNA is more stable than in conventional amplification methods. This attribute makes the DNA extremely susceptible to contaminating surfaces and future reactions if the samples, after amplification, are not properly contained. “Never open the tubes after an amplification” is a mantra when working with LAMP haha.
Having said that, there are new variations of LAMP that help avoid false positives. Here are some of my favorites for real-time LAMP:
-> Molecular beacons: https://genomebiology.biomedcentral.com/articles/10.1186/s13059-021-02387-y
-> DARQ: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7860930/
They employ the concept that regardless of the DNA amplified in the tube, the signal will only be generated if that DNA is the specific one you’re looking for. If the amplification is just noise the signal generation mechanism won’t be triggered.
(Sorry for the duplicate comments, the webpage took 1 hour to update them so i thought they were getting filtered by the spam detection algorithm)
For IVDR based diagnostic devices PPV and PNV % are demonstrated with comprehensive analytical and clinical studies. Amplification in black samples is true and also controlled for using LOB and incorporated into the uncertainty of results. These amplification technologies can provide solid diagnostics when CE IVDR.
The lying by the CPC about every detail while shutting down incoming traffic and keeping outgoing flights open was criminal and the kind of thing seen as an opportunity to take advantage of. Followed by US and EU “experts” trying to explain how the first three deaths from the Chinese Bat Fever were the top three researchers at the Chinese Bat Fever lab did not mean it got loose form the Bat Fever laboratory – which got its funding through the American “expert”.
The whole thing has been a massive clusterf*ck of government over-reach. Aside from total isolation, the only thing that really worked was hand washing and don’t touch your eyes or nose (or face in general) before you wash your hands. Did vaccines make a big difference? Probably not a lot, especially the Chinese vax. These viruses always decline in virility over time. If they are too aggressive they incapacitate the host before they can spread. Ask yourself why every new flu dissipates in less than a year? I wonder how many adult American deaths were obese?
It sound like you don’t have any personal experience with friends or loved ones dying from SARS infections. 8 friends of mine died from COVID, some quickly from clots, and some slowly from massive and irreversible lung fibrosis. The worst one was the dad of a couple I married. He was a father with 5 kids. They were afraid to get vaccinated and they all ended up getting COVID at a large family gathering. He started feeling sick, and a couple of days later ended up in the hospital. He died in the ICU after about a week.
Anecdotes need a lot more info than that. There were actually a few children who dies. Some with cystic fibrosis, some with lupus and immune suppressing drugs. Some in chemo from cancer. Some morbidly obese. I don’t know of any who were going to public school but one would think so. Just look at the data https://www.cdc.gov/nchs/nvss/vsrr/covid_weekly/
Death causes weren’t properly evaluated. Anyone who was tested positive and died soon after was counted as “died of corona”, even fatal accidents. It took more than a year for the official wording to slowly change to “died with corona”. Also we don’t know how many deaths were caused by misdiagnosis and improper treatment, like undiagnosed pneumococcus infections. Or from getting no treatment at all, as many hospitals delayed planned treatments. A serious quality control of vaccination, tests, treatment and protective gear was completely missing. Instead, anyone expressing his doubts about that was instantly defamed as a right-wing weirdo.
The only positive side effect is that the mass production of PCR tests and mRNA vaccines made them darn cheap, which, if used wisely, can be benficial to mankind, but they should be kept out of reach of politicians and other self-proclaimed Messiahs.
That’s not correct. A series of objective criteria and a sequence of events that are needed by the coroner on the death certificate to indicate a primary or secondary cause of death from COVID. These diagnostic criteria remained consistent during the pandemic.
I can’t even. But to say a PCR is out of range for hospital scale diagnostics is … no. Just. No.
Is this the new LAMP? Will it run a server?
qLAMP. Probably only runs on quantum computers.
Fascinating. Besides the obvious (deaths, long COVID), the pandemic seems to have caused a kind of cognitive disfunction.
OTOH, there is historical precedent: in Western Europe, the Black Death 1350 triggered quite a few such irrational things, too.
Thank you so much for featuring our work! It’s incredibly gratifying to wake up and see this on a platform I’ve always admired!
I apologize that the photos and the GitLab repository are a bit outdated. Over the past few months, we’ve been rushing to validate some experiments aimed at repurposing this hardware for other biological protocols, such as synthetic biology and cell-free experiments, characterizing locally produced enzymes (their activity and stability), and studying bacteria interaction and viability. We have an open-source publication featuring all these protocols and results coming soon!
I’m currently updating the repository, but in the meantime, here is an updated image of the hardware: https://gitlab.com/open-bioeconomy-lab/diagnostics-hardware/rt-lamp-device/-/blob/master/Photos/new_electronics.png
Have a great day, and thank you again for the feature! For those of us starting in open hardware, it means a great deal to have the opportunity to see our work featured on this platform.
Bonus image, an electronic diagram of all the modules and a simplify view of the working principle of the qLAMP: https://gitlab.com/open-bioeconomy-lab/diagnostics-hardware/rt-lamp-device/-/blob/master/Photos/diagram.png
Thank you so much for this project! One question… How much possible do you find in the future, of open source DNA sequencing hardware? Maybe it’s unfeasible due to nanopore technology? Your take on it?
it seems like this needs a supply of reagents specialized to the target it’s testing for?
pretty neat but in star trek Dr Crusher can test for *novel* nucleic acid sequences with a handheld gadget, without sending back to the lab to get a special testing serum for each new disease she discovers :)
Hahaha!! It needs special “primers” (small sequences of dna that nucleate the reaction specifically for the target you are looking to). They are kind of easy to develop (if someone haven’t develop something already) and they cost around 50€ for hundred of reactions. For the rest of the reagents (polymerase and salts) there are folks as the open bioeconomy lab (https://openbioeconomy.org/) trying to develop open source protocols to allow local producers to make them locally.