Sporal Test — Biological Autoclave Verification Step by Step [2026]

A sporal test is a biological sterilisation indicator -- the only method that gives a definitive answer to the question: did your autoclave actually kill all microorganisms, including bacterial spores? Chemical indicators that change colour only confirm that a process took place, not that sterilisation was successful. Sanepid (Poland's State Sanitary Inspectorate) requires a biological test at least once a month, and the result must be documented. If you are not yet sure how to do this correctly, this article will walk you through the entire process step by step.

What a Sporal Test Is and Why It Is Mandatory

A sporal test (biological test, biological sterilisation indicator) is a preparation containing spores of Geobacillus stearothermophilus bacteria. These organisms are exceptionally resistant to high temperature and moisture -- if your autoclave can destroy them, it will destroy all other pathogens as well. That is why the sporal test is the gold standard for verifying sterilisation effectiveness.

The legal basis is the Regulation of the Minister of Health on sanitary requirements for premises and equipment of hairdressing, cosmetic, tattoo and wellness establishments (Journal of Laws 2002 No. 190, item 1591, as amended). During inspections, Sanepid checks the autoclave control log and may request biological test results for the past 12 months. Missing documentation is the most common reason beauty salons receive fines.

An important distinction: chemical indicators of classes 4, 5 and 6 are process indicators -- they confirm that an instrument was exposed to the appropriate temperature and time, but they do not confirm sterility. A biological test is the only sterility indicator.

How to Perform a Sporal Test Correctly

There are two types of biological tests available on the market: ampoules sent to an external laboratory (results within 24-72 hours), and self-contained biological indicators (SCBIs), where you carry out the culture yourself using a dedicated incubator after 24 or 48 hours.

Step-by-step procedure:

1. Place the biological indicator in the coldest spot of the autoclave chamber -- usually near the drainage outlet, at the lower rear of the chamber. This is the place where steam arrives last and where the temperature is lowest.
2. Place the test alongside a normal instrument load -- do not run a separate cycle just for the test.
3. Run a full sterilisation cycle according to the parameters for that load (e.g. 134°C for 18 minutes for unwrapped instruments, or 134°C for 5 minutes for wrapped ones).
4. After the cycle ends, remove the indicator and activate it according to the manufacturer's instructions (crushing the ampoule with growth medium in the case of SCBIs).
5. Place in the incubator for 24-48 hours at the temperature recommended by the manufacturer (usually 55-60°C), or send it to a laboratory.
6. Read the result: no colour change in the medium = negative result = sterilisation successful. Colour change (usually from purple to yellow) = positive result = FAILURE.

The cost of a single biological test ranges from 30 to 80 PLN depending on the manufacturer and method (SCBIs are more expensive but give results faster and without the need to send anything by post).

What to Do with a Positive Result -- the Failure Protocol

A positive biological test result is a serious situation requiring immediate action. It means your autoclave was not sterilising effectively -- at least in that one cycle, but probably for a longer period.

Action protocol:

1. Stop using the autoclave immediately. No instrument may leave the device labelled as "sterile" until the fault has been investigated.
2. Identify and recall all instruments sterilised since the last negative test. If you test monthly, this could mean instruments from an entire month -- all of them must be re-sterilised in another working autoclave or destroyed.
3. Contact the autoclave service department or manufacturer. Check: chamber seals, filter, temperature and pressure sensor, safety valve. Request a service report.
4. Document the entire incident in the control log: date of failure, cycle number, actions taken, repair date and date of retest.
5. Carry out at least 3 further biological tests after the repair to confirm the device is working correctly.
6. If you served clients during that time -- you do not need to notify them if instruments were single-use or sterilised in a verified alternative autoclave. If that was not the case, consult a lawyer.

Documentation -- What to Enter in the Log

Every biological test must be recorded in the autoclave control log. This log is an official document that you can present during a Sanepid inspection.

Each entry should include: test date, autoclave serial number, cycle number/identifier, cycle parameters (temperature, time, pressure), type and batch number of the biological indicator, result (negative/positive), and the signature of the responsible person. If the result was positive -- add a description of corrective actions taken and the date of the retest with its result.

Documentation must be kept for a minimum of 5 years. It is worth photographing indicators (both negative and positive) and archiving the photos together with the log entry -- in case of any dispute.

Frequently Asked Questions (FAQ)

Do I need to perform a sporal test every month, or more frequently?

Sanepid requires a minimum of once a month. Autoclave manufacturers and European standards (EN 13060) recommend a biological test after every repair, after a long break in use, and after any change to the sterilisation programme. Many specialists in busy salons run the test weekly.

Do I need to send the test to a laboratory, or can I do it myself?

You can use self-contained biological indicators (SCBIs) with your own incubator -- you read the result yourself after 24-48 hours. You do not need to send anything to a laboratory. It is important that the incubator is calibrated and that you follow the indicator manufacturer's instructions precisely.

What is a class 5 chemical indicator and does it replace a biological test?

It does not replace it. A class 5 indicator (integrator) changes colour when all critical process parameters have been reached (temperature, time, saturated steam). It is an excellent process indicator -- but it indicates the "process", not "sterility". The only proof of sterility is a biological test.

How much does an incubator for biological tests cost?

Incubators for SCBI tests cost from 500 to 2,000 PLN. This is a one-off cost that pays for itself within a few months (you save on laboratory postage). Before buying an incubator, make sure it is compatible with the specific brand of indicators you intend to use.

Can a Sanepid inspection officer take away my autoclave control log?

An inspector may review the log on site and make copies, but should not take the original without a formal decision. For safety, keep the log in electronic format or make regular copies of the pages -- in case of any misunderstanding.