Enterovirus antibody neutralization test in Europe

[green_echo]

Hello,

I've been trying to get the coxsackievirus and echovirus antibody neutralization tests done for some time now with no luck. If anyone has any knowledge of how to get these tests done in Europe now, I'd really appreciate it! I am based in the UK but happy to travel. Also open to FedExing samples to the US if that's something anyone has tried.

Here's what I've tried so far, per the (wonderful!) ME/CFS road map.

• Frankfurt University Hospital, Germany - the university was hit with a cyberattack last year so most of the links and forms no longer work. I have only managed to speak to someone who works in the invoicing department at the hospital, who didn't speak much English. Has anyone managed to get the test done here?
• University Hospital Erlangen, Germany - they said they no longer offer the coxsackievirus test
• Torlak Institute of Virology, Serbia - currently trying to get them to accept a sample if I can FedEx it - will update here if successful
• Hellenic Pasteur Institute, Greece - no response yet

If anyone has any ideas of how to move forward, I would really appreciate it! Thank you

 

[Cipher]

Frankfurt University Hospital, Germany - the university was hit with a cyberattack last year so most of the links and forms no longer work. I have only managed to speak to someone who works in the invoicing department at the hospital, who didn't speak much English. Has anyone managed to get the test done here?

I've tested via them, see this post.

 

[Canned]

I have done the testing in Frankfurt in 2021. i would like to check my titers but couldnt find a link that works. Is it just their website not working or is their Service no longer avaiable? I might call them next week

 

[green_echo]

I've tested via them, see this post.

Thanks so much, so just to confirm

• One 5ml sample is needed for each test
• The samples need to be centrifuged
• You send the sample directly to the virology department at the hospital with the form enclosed

And is this form the correct one? I can only find it on WayBack machine.

Thank you!

 

[green_echo]

I have done the testing in Frankfurt in 2021. i would like to check my titers but couldnt find a link that works. Is it just their website not working or is their Service no longer avaiable? I might call them next week

This is the contact info on the website:

Secretariat Prof. Ciesek:
Tel. 069 6301-5219
Fax 069 6301-6477
Email Virologie@ukffm.de

Operating hours:
Mon + Wed 8:00 a.m. - 1:00 p.m

Please do let me know if you get any more info

 

[Cipher]

Thanks so much, so just to confirm

• One 5ml sample is needed for each test
• The samples need to be centrifuged
• You send the sample directly to the virology department at the hospital with the form enclosed

And is this form the correct one? I can only find it on WayBack machine.

Thank you!

I found an old e-mail from them stating that 1 ml serum is enough. I don't know if they meant 1 ml serum per panel or 1 ml serum in total.

A blood sample is drawn into a serum separation tube, which is then centrifuged, from which the serum is pipetted into a transport tube without additives.

On my computer I found the form I used, which seems to be a slightly newer version than the one on the Wayback Machine. I've attached it to this post.

I filled it out like this (you'll have to translate and double-check):

Spoiler

I put my e-mail address at the bottom even though it says telephone, and I got the results via e-mail.

I sent it to the postal address listed at the top left of the form.

I don't know if the cyberattack have affected their ability to send e-mail results, but I guess they will fall back to regular post if they have issues.

I included my country in the address fields, but if I remember correctly the invoice letter I received missed the country name anyway. However, I wrote the postal code with the country prefix as a backup. Since you're in the UK I think it is GB-XXX XXX. That way the postal service knows what the destination country is even if they have omitted the country line. They could probably figure it out without the prefix also, but it might delay it.

 

[Hip]

University Hospital Erlangen, Germany - they said they no longer offer the coxsackievirus test

Thanks for this info, I will remove it from the roadmap.

The samples need to be centrifuged

For antibody testing, labs need the blood serum, rather than the whole blood. The serum is the pale yellow liquid obtained by centrifuging whole blood to remove the blood cells.

If you tell them you need serum, a phlebotomy lab which draws your blood will centrifuge there and then (takes about 20 or 30 minutes to centrifuge), and hand you a test tube of your serum, which you can then courier to the lab for antibody testing.

 

[green_echo]

Great, thanks very much! I will update if I manage to do the test successfully

 

[green_echo]

Update: Despite my best efforts, the samples ended up stranded in German customs and eventually would have spoiled, so I asked for them to be destroyed. Outside of the customs union (I'm in the UK), it may be too difficult to send them over.

 

[Cipher]

Update: Despite my best efforts, the samples ended up stranded in German customs and eventually would have spoiled, so I asked for them to be destroyed. Outside of the customs union (I'm in the UK), it may be too difficult to send them over.

I'm sorry to hear that. Did they tell you why they held the package? What shipping company did you use and how did the customs declaration look?

 

[green_echo]

Hey sorry for the delay! They said they needed more info from the recipient and I couldn't get hold of the lab. One weird thing was that the carrier I used -DHL- wouldn't let me put exempt human specimen on the box. So perhaps if I'd had that it would be OK. I used a free online website to do the customs declaration but can't seem to find it now.

I did end up going in person to Frankfurt and dropping a sample off myself, which they say they have received. It should take about 2 weeks. The website is up again after the cyberattack and is much easier to use.

I called and spoke to someone on +49 69 6301 5219 to confirm receipt of the sample and the person spoke very good English, if that helps anyone.

 

[MartinK]

Guys @green_echo @Cipher I want to test the entire list of enteroviruses in Frankfurt because our Czech laboratory failed and no longer offers services to the public.
- please, what was the total amount of serum you sent?
- does the centrifuged serum have to be separated to another tubes?
- time limit for the shipment to reach the laboratory?
- did the address have to be specified, or is the one in the upper left corner of the request sufficient?

Thank you for help!

*By the way, has anyone considered buying a centrifuge for home use?
Like this: https://www.kaufland.cz/product/328...aid&utm_campaign=pricecomparison&sid=22773832

 

[Cipher]

- please, what was the total amount of serum you sent?

I don't remember exactly what I sent, but see below:

I found an old e-mail from them stating that 1 ml serum is enough. I don't know if they meant 1 ml serum per panel or 1 ml serum in total.

- does the centrifuged serum have to be separated to another tubes?

Some types of serum separation tubes contains a gel that keeps the blood cells separated from the serum even during transport, which removes the need to pipette the serum into a separate tube. However, these tubes are supposed to be used in a swing out centrifuge, and are not recommended to be used with a fixed angle centrifuge due to the gel seal becoming weaker.


My local phlebotomy clinic refused to draw and centrifuge my blood for external labs, so I did it at home. I used a red-cap serum tube without a gel, and then pipetted the serum to a white-cap non-additive tube.

- time limit for the shipment to reach the laboratory?

ARUP says that it's stable for at least 48 hours in ambient temperature.

- did the address have to be specified, or is the one in the upper left corner of the request sufficient?

I sent my serum to the address listed at the upper left corner of the request form.

*By the way, has anyone considered buying a centrifuge for home use?
Like this: https://www.kaufland.cz/product/328...aid&utm_campaign=pricecomparison&sid=22773832

I bought a ~60 USD centrifuge that looks like this:



It's of pretty bad quality, but works. It heats up the samples too much if used as intended, however this can be worked around by removing the lid and some of the tubes to allow air into the machine. It starts to shake at some speeds, and printed rpm is not accurate. I used a laser tachometer to manually measure the rpm and calculate the g-forces. It's important to weight-balance a centrifuge, or it will break.

Here are some bookmarks that I have saved that might be helpful:

Spoiler: Links

https://www.gbo.com/fileadmin/media...der_of_Draw_en_rev03_2021-04_DIN-A1_print.pdf
https://web.archive.org/web/2022011...980200_IFU_VenousBloodCollection_rev23_EN.pdf
https://www.thepathologycentre.org/...1.UN-Griener-tube-chart-and-order-of-draw.pdf
https://druckerdiagnostics.com/wp-content/uploads/2017/09/Greiner-Centrifugation.pdf
https://www.gbo.com/fileadmin/media...tion_Techniques_booklet_en_rev08_0122_web.pdf
https://www.labcorp.com/resource/blood-specimens-chemistry-and-hematology
https://www.wolframalpha.com/widgets/view.jsp?id=1192007d993f13fc9de7ce32bd9bab16
https://www.youtube.com/results?search_query=butterfly+needle

 

[MartinK]

@Cipher Hey, thank you for this informations!
Swing out centrifuge will be expensive I suppose... I tried to find some, but the chat gpt was quite lying.

I have some serum tubes with gel at home. I can't find out if other types of test tubes can be purchased in smaller quantities in Europe, because I always only find large packages of hundreds of pieces.

I would rather send them 5 ml of serum to the Frankfurt laboratory, I believe that will be more certain.

 

[MartinK]

@Cipher did you really sended centrifuged sample? Im super confused about their instructions:

first mail:
For this we need a large white serum tube (10 ml) . non-centrifuged serum. The tube must not contain any additives.

second mail:
I apologize - that was perhaps a little unfortunate. It must be serum. Not whole blood. – From the Serum 2 ml is enough

So, how you understand it? - taking blood draw (2 ml) straight into a white tube without additives? No centrifugation/separation is needed? (by the term "serum" I understand what is created from the blood after centrifugation)

 

[Cipher]

@Cipher did you really sended centrifuged sample

Yep, I sent serum that I had pipetted from a centrifuged serum tube to a white non additive tube.

I interpret those emails as they want 2 ml of serum in a non additive tube. To generate 2 ml of serum one needs to draw approximately 2.5 times the amount of blood, i.e. 5 ml. It's always a good idea to draw too much rather than too little. It becomes easier to pipette also if you have some spare, so that you don't accidentally pipette some blood cells.

 

[Inez]

@Cipher did you really sended centrifuged sample? Im super confused about their instructions:

first mail:
For this we need a large white serum tube (10 ml) . non-centrifuged serum. The tube must not contain any additives.

second mail:
I apologize - that was perhaps a little unfortunate. It must be serum. Not whole blood. – From the Serum 2 ml is enough

So, how you understand it? - taking blood draw (2 ml) straight into a white tube without additives? No centrifugation/separation is needed? (by the term "serum" I understand what is created from the blood after centrifugation)

I had exactly the same confusing info over two emails. I'm going to email again because I now have no idea if the serum should be centrifuged or not? If blood is left to stand for a certain time it separates into serum right? I don't know if they mean that.

 

[Cipher]

I had exactly the same confusing info over two emails. I'm going to email again because I now have no idea if the serum should be centrifuged or not? If blood is left to stand for a certain time it separates into serum right? I don't know if they mean that.

While you technically can skip the centrifugation and wait hours for the blood cells to separate by gravity, this is not a method that is used when it comes to blood analysis:

if the whole is allowed for a long period, it will be separated. However, the separation will not be appropriate when compared with samples that were separated using a centrifuge. Besides, the time is usually very short using a centrifuge compared with when allowed to stand.

https://www.researchgate.net/post/T...d_from_serum_or_plasma_without_centrifugation

NOTE: Tubes should be centrifuged no later than 2 hours after collection. Extended contact of blood cells with the serum or plasma, may lead to erroneous analysis results, hence centrifugation might be necessary sooner depending on the analyte. It is not recommended to re-centrifuge gel tubes once the barrier has been formed. The debris underneath the gel might contaminate the supernatant.

https://web.archive.org/web/2022011...980200_IFU_VenousBloodCollection_rev23_EN.pdf

The "non-centrifuged serum" must be a miswrite in my opinion.

 

[MartinK]

From one German ME/CFS friend:

"I sent whole blood but within a serum tube, so that they can centrifuge it"

Guys, now Im totally confused!

 

[Cipher]

From one German ME/CFS friend:

"I sent whole blood but within a serum tube, so that they can centrifuge it"

Guys, now Im totally confused!

Technically possible, but is not a normal way to do it, which is why they wrote "Not whole blood" in the email.

See my previous post, the second quote is from the instruction manual for Vacuette vacutainers.