Simultaneous quantification of thyroid hormones using an ultrasensitive single-molecule fourplex nanoimmunosensor in an evanescent field

[helvella]

If this sensor lives up to the claims and is inexpensive, we could be on the cusp of major changes in thyroid testing.

Seems obvious that once you are going in the direction of single-chip analysers, someone will be looking at incorporating it into a mobile phone.

(And no, I don't understand how they are doing this!)

Biosens Bioelectron

. 2023 Jan 15;220:114894.

doi: 10.1016/j.bios.2022.114894. Epub 2022 Nov 7.

Simultaneous quantification of thyroid hormones using an ultrasensitive single-molecule fourplex nanoimmunosensor in an evanescent field

Junghwa Lee 1 , Seungah Lee 2 , Gwang Lee 3 , Seong Ho Kang 4

Affiliations

Affiliations

• 1 Department of Chemistry, Graduate School, Kyung Hee University, Yongin-si, Gyeonggi-do, 17104, Republic of Korea.

• 2 Department of Applied Chemistry and Institute of Natural Sciences, Kyung Hee University, Yongin-si, Gyeonggi-do, 17104, Republic of Korea.

• 3 Department of Physiology, Ajou University School of Medicine, 164, World cup-ro, Yeongtong gu, Suwon si, Gyeonggi do 16499, Republic of Korea; Department of Molecular Science and Technology, Ajou University, 164, World cup-ro, Yeongtong gu, Suwon si, Gyeonggi do 16499, Republic of Korea.

• 4 Department of Chemistry, Graduate School, Kyung Hee University, Yongin-si, Gyeonggi-do, 17104, Republic of Korea; Department of Applied Chemistry and Institute of Natural Sciences, Kyung Hee University, Yongin-si, Gyeonggi-do, 17104, Republic of Korea. Electronic address: [e-mail].

• PMID: 36371958

• DOI: 10.1016/j.bios.2022.114894

Abstract

The thyroid gland, which regulates the metabolism of the human body, has a sophisticated feedback system that induces the secretion of thyroid-stimulating hormone (TSH) to regulate the levels of triiodothyronine (T3) and thyroxine (T4). In this study, a single-molecule fourplex nanoimmunosensor was developed for the simultaneous quantitative analysis of TSH, T3, and T4. The three thyroid hormones were detected with a high signal-to-noise ratio in an evanescent field using laser-induced total internal reflection fluorescence. Additionally, the use of gold nanoislands for the detection of molecular interactions between thyroid hormones and antibodies labeled with quantum dots minimized the background noise from the substrate compared with the use of microislands or microwells. The nanoimmunosensor exhibited excellent detection limits of 114-193 yM (yoctomolar = 10-24 M) for thyroid hormones. The detection sensitivity was approximately 1015-fold higher than that of the conventional enzyme-linked immunosorbent assay. Paired Student's t-test of the human blood samples revealed that the difference between the two methods was insignificant at the 98% confidence level. Therefore, the proposed single-molecule fourplex nanoimmunosensor can be used for early diagnosis and prognosis monitoring at the single-molecule level because it can accurately, rapidly, and simultaneously diagnose various thyroid diseases, such as hyperthyroidism and hypothyroidism.

Keywords: Nanobiosensor; Quantum dot; Simultaneous quantification; Single-molecule fourplex nanoimmunosensor; Thyroid hormones; Yoctomolar detection.

Only abstract available:

pubmed.ncbi.nlm.nih.gov/363...

sciencedirect.com/science/a...

[humanbean]

In this study, a single-molecule fourplex nanoimmunosensor was developed for the simultaneous quantitative analysis of TSH, T3, and T4. The three thyroid hormones were detected with a high signal-to-noise ratio in an evanescent field using laser-induced total internal reflection fluorescence.

Are they referring to Total or Free levels of T4 and T3.

And for people who appear to be very clever, referring to TSH as a thyroid hormone is not impressive.

[helvella in reply to humanbean]

I agree - and, without the full paper, it is not clear whether they are Total or Free tests.

However, for early testing, it does have promise. Might be possible to convert a Total test into a Free test once you get it all working well.

Completely agree about TSH. But I suspect the train of thought is that all three are often considered as part of the Thyroid Function Test/Thyroid Panel. Poor language. But better than my Korean.

[Tythrop in reply to humanbean]

Yes to the point about tsh .it is as much a Thyroid hormone as the oil warning light on my car dashboard is to what the oil does in the engine . This is very relevant at present as I'm on the cusp of another attempt to get T4 from GP but I'm not expecting it as , tsh is showing on bloods as about 25% above bottom of range ,BUT my T4 result is 8 when the bottom of the range is 12 ..that's 4 less than the minimum NHS range protoco.The TSH IS like the car's warning light not showing when the engine oil dip stick says it is well below minimum but the garage/ GP saying I don't need oil when infact its a dodgy oil warning light .

[humanbean in reply to Tythrop]

That sounds like you have Central Hypothyroidism. In other words your thyroid may be perfectly healthy but your pituitary is not capable of producing sufficient TSH to stimulate your thyroid to do its job of producing all your T4 and some T3.

[jimh111]

First time I've seen the prefix 'yocto' used in a study. Quite impressive 'Startrek' stuff. What happens when they misplace this tiny sensor?

(click on the magnifier to see the bottom of the table).

x

[helvella in reply to jimh111]

First time I've ever posted about yocto!

[tattybogle in reply to helvella]

is yocto a tiny mole ?

tiny mole

[turquoisea7 in reply to tattybogle]

I love moles

[helvella in reply to turquoisea7]

Many years ago, on a summer evening, I was at a pub with a garden by the river. Nothing special - just a friendly local.

As it was warm, the french doors were open. And somehow, I really don't know exactly how it happened, a mole from the garden went inside. Being a mole and surrounded by people - it was in a panic and did what a mole does. It tried to dig - and managed to get under the carpet.

You could see a small mound whizzing all over the place. Phenomenally fast.

Eventually it was persuaded to get to the edge near the door and escaped. Poor thing.

[tattybogle in reply to helvella]

When i lived in yurt we didn't have a wooden floor like the posh glampers of today , we had an insulated groundsheet/ layers of carpet . The woodburner in the middle was on 1 inch raised metal hearth plate.... one spring when we took everything down to move , there was an amazing pattern of mole activity radiating out from the centre under the burner .... they'd obviously been enjoying 'Overhead Heating at Mole Head Quarters' ... still makes me smile to think of what our moles were saying to thier mates inthe pub ...."Everyone back to 'ours' .. the ceiling is warm "

[helvella in reply to tattybogle]

Do you get moles in where-yurts-come-from?

[tattybogle in reply to helvella]

apparently so .. an Altai Mole en.wikipedia.org/wiki/Altai....

[tattybogle]

Dear Santa . i have been very good , please can i have a a single-molecule fourplex nanoimmunosensor on a gold nanoisland for Christmas ?

[TiggerMe Ambassador in reply to tattybogle]

Yes dear.... would you like the Russian, Chinese or Korean one 😏

[tattybogle in reply to TiggerMe]

Mmm ...tricky question ...

[Aurealis]

Maybe I am the only one whose heart sank when I read this. My worry - it’s more “scientific”, it’ll be much harder to argue back to the “you have to reduce your dose” brigade, when they’re armed with such high tech science.

It’s measuring t3,t4 and TSH in blood again isn’t it? Whereas it’s cellular levels that make the difference.

[1tuppence in reply to Aurealis]

And aren't we all individuals who react in our own individual ways to whatever we're living with/exposed to?

[helvella in reply to Aurealis]

It does have several potential positives:

More accurate. Current tests are inconsistent. Hopefully no new test will be approved without being substantially more consistent.

Cheaper (hopefully) so frequent tests can be done.

All three on one device so we should expect no more "Only done TSH" disappointments.

If it can be done on/with mobile phone will be under patient control. For freedom of timing, frequency, etc.

None of this waiting six weeks for these to stabilise. Just test every day, even several times a day, and see how it changes over time. If it looks like it is going to end up short, consider incrementing (slightly) early. If it looks like it is going to overshoot, drop (slightly) when you realise.

Use sophisticated software to interpret results.

If a medicine batch is sub-potent (or super-potent), of changing make has an impact, find out more or less as it happens. Not weeks, months, even years later.

I suspect we'll never have in-cell T3 and T4 testing. We need to look at what T3 and T4 do and look for those effects rather than the actual hormones.

Science and scientific method aren't really the baddies. It is misunderstanding and misapplication of them.

[TiggerMe Ambassador in reply to helvella]

Yes, the pro's definitely out weigh the con's, it seems eminently sensible to be able to take readings as diabetics do... how empowering! 👏

[Zephyrbear in reply to helvella]

“Hopefully no new test will be approved without being substantially more consistent”

When did consistency ever play a part in testing when money is at the bottom of it all? Forgive my cynicism, but I don’t believe that the best interests of the patients are ever uppermost in considerations for anything “new”… especially where testing and big pharma are involved.

Good if it happens, but I’m not holding my breath.

[helvella in reply to Zephyrbear]

In some areas, there has been a fairly successful harmonisation process. For example, I think basic blood count tests are reasonably consistent across analysers.

But far too many aren't.

[Zephyrbear in reply to helvella]

Which may be the reason why, in the “good old days of yore” when doctors actually spent time observing and listening to patients, they were rather more successful at diagnosing things like hypothyroidism earlier instead of having to wait for a TSH reading on a blood test to reach a certain magic number before treating said patient…

[Beads in reply to Aurealis]

It’s measuring t3,t4 and TSH in blood again isn’t it? 

But the difference here is that it will measure all 3, rather than just the TSH that the doc tests. We’ll be able to see the interactions between TSH and actual thyroid hormones more regularly; this might mean less dependence on TSH being the be all and end all.

[Aurealis]

agree with all of that 👍

[waveylines]

Can't we get away from blood tests? This over reliance on them drives me mad! They are a guide - nothing else. This idea that a blood test tells all is an illusion but one that big pharma love. Lazy doctoring. Blood tests are helpful when used with a full medical evaluation which involves looking at and talking to the patient. Sorry to be a kill joy but this strikes me as more of the same with a few tweakes thrown in....lol

[TiggerMe Ambassador in reply to waveylines]

It's got to be useful to prove that if you perhaps don't fit the 'norm' and your optimal falls outside the current ranges they so love, you can at least personalise it and monitor/ pinpoint more accurately what suits your needs?

I like the idea of fine tuning rather than the rather hit and miss we current have

[helvella in reply to waveylines]

Nonetheless, still better than poorer tests done less frequently.

Have a look here:

healthunlocked.com/thyroidu...

[waveylines in reply to helvella]

Sorry been in for an operation....Im lucky as it went ahead even though Christmas this year I will be convalescing.

I hear you, Helvella, but what worries me is the lack of looking at symptoms and signs. The physical examination has gone.... And what has replaced it isn't that great leaving people feeling soo poorly. I had an old school doctor - he could tell I had hypothyroidism the minute I walked through the door. I had the classic moon face, puffy dull eyes & my general.presentation was slow. He looked at my skin under a magnifier, looked at my hair, eyebrows, eyes, took my pulse and blood pressure etc... He listened & heard my dlow speech, throaty voice, obvious loosing of words and so forth. He knew his stuff. A blood test showed I was grossly under treated but my GP hadn't the where within to spot that. GP just said you are fine! Bull!! I could barely walk & was struggling to work & was in a lot of pain. I was already on treatment and diagnosed with Hypothyroidism & the blood test showed TSH 5.6, Ft4 10 (range back then. went up to 23. ) Medics AND patients need to be educated to look at signs and symptoms on top of blood tests....& how to check a patient physically for hypothyroidism.

Am grateful to the doctor I saw. His old school spproach saved my life and though it took time he restored my health.

These days you can look grossly hypothyroid but if your TSH isn't over range of 10 plenty of GPs will say you are fine......even though if they used their eyes it would be obvious that this isn't the case. So I take blood tests with a very large pinch of salt.......they're an indicator. I dont believe anything can truly replace a proper skilled examination from a knowledgeable doctor. Blood tests are there to support not replace. They are generic & have led to disengagement from the patient, which has led to poor testing protocols like treating by the TSH! It's a rat hole that costs the NHS money and has NOT led to more effective treatment for many.....quite the opposite.

[tattybogle in reply to waveylines]

i agree , there is a desperate need to improve awareness in GP's of actual signs and symptoms/ presentation of hypo / hyper / overmedication/ undermedication.

i saw 2 GP's and a nurse....not one of whom thought to consider my thyroid status or re check my TFT's . between them they sent me for kidney US, FBC ,urine sample, endoscopy . colonoscopy and CT scan . The actual problem was slight overmedication with Levo , but it took an elderly semi retired GP to diagnosed it ...( by looking, asking the right questions ,listening to the answers and asking me to hold out my hands) . I saw her purely by chance while chasing up a 2 week referral for one of the above tests.

The other 3 (without her knowledge and experience of how overmedication actually presents) , either didn't consider it at all , or perhaps relied on my 10 month old 'annual TFT review' looking fairly 'normal for me' .. and consequently wasted a huge amount of time and NHS resources. ,, not to mention giving me an unnecessary dose of iodine contrast/ CT scan .

MT TFT was pretty unremarkable .. the fT4 was a bit over @120%. but it had been higher than that several times before without causing any symptoms , and my TSH was 0.018 instead of my usual 0.05 ... so the observation of my whole presentation by an experienced GP was the key here , and was much more use than the blood test in isolation.

wishing you a 'Happy Convalescence' Waveylines , x

[helvella in reply to waveylines]

Wishing you well in recovery.

Definitely agree that examination and observation has fallen away.

Can't help thinking that we need to look at multiple factors - and some are observations (e.g. how person walks in, sits down, behaves), some examinations, and some are tests. Combining all of them is necessary to achieve near-100% diagnosis and best choice of treatment.

[TiggerMe Ambassador]

Is it likely we could be treated more like diabetics?

I'm intrigued how it works in other countries where they can just go to the pharmacist?

[tattybogle]

anyone paying attention may have noted i'm a confirmed luddite. But .... a device that could tell me my TSH / fT4 / fT3 whenever i wanted to ask it ? ... this ... i do want.... not so that Doctors can see the results ... (i'm sure many of them would still find ways to misuse the information) ... but for myself , so i could learn more about me , by my own observations and record keeping .

i agree, a method of seeing cellular levels is unlikely outside of labs .... but by being able to observe our own T4/T3 /TSH changes in real time we would be able to see and hopefully get to understand so much more about 'what is going on within' that we can at present .

[helvella in reply to tattybogle]

I also feel it likely that if we could, for example, do a fingerprick test (and maybe only need a single drop) at will, that would be advantageous to us.

We might start to really find the levels at which we feel best. (Ever tried to ask a GP for a test because you feel well?)

We can check if we feel wrong - including if we have changed the medicine we are taking. Picking up subtle changes over extended periods like months and years.

[waveylines]

That's true Helvella. I actually think the treatment we take is quite crude. We have to allow for the cold the heat, if we are unwell, stressful time, excessively busy times etc. We are prescribed a fi ite daily amount.....it does not allow for natural variations as above that a working thyrois would adjust accordingly. I well remember a well known esteemed medic who helped a lot of patients with hypothyroidism saying he takes a bit extra on a busy day. Says it all. I dont think you will ever find a definitive amount that does all. Therefore its really important we are vigilant on signs symptoms of over and under for each person. These can vary from person yo person. I'm very stable on my dose and have been for years now....mainly because my thyroid is well and truly dead! Even so I have to do tweaks up and down. I can tell when I'm a tadge over and equally the other way. I don't need a blood test for this. Only if I need reassurance that I'm sticking in range for my thyroid hormones. I remember having good blood thyroid levels.....I felt rubbish. What was wrong was I wasn't addressing the cofactors. Once I did that & was on a ndt....boom!

Diabectics have simular problems but they do have those tracker systems now which must be marvelous. Loved to know what they really think of them or are we just bring taken in by the media hype?

No chance with us. We are still at a basic level, getting doctors to recognise the importance of thyroid hormones and the need to test ft3 as well as ft4, and in a fair few cases to test beyond the TSH! 🙈

Its a long way to Tipperary, long way to go....🎵🎶

[helvella in reply to waveylines]

I know of someone locally who has a diabetes device - and finds it wonderful.

When you are an old hand (not being rude!), I think making adjustments is not so difficult. But starting out, it is devilish. And so easy to take a tiny bit more for a few days, then find you are over-dosed. Or just not be sure.

[TiggerMe Ambassador in reply to helvella]

Something similar has to be the future and would allow self management

[helvella in reply to TiggerMe]

Which is very much what I'd like to see.

(With support, as needed, of course.)