Methodology – Saliva Hormone Testing

Saliva Hormone Testing
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Saliva Testing Methodology
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Direct Assay vs. Extraction Process

At Labrix it is our goal to set the standard for the most accurate and reliable salivary hormone testing. We employ the most stringent methodology available to ensure that when you test through our laboratory your results are accurate. Unfortunately, this level of testing does not always occur at other salivary hormone testing laboratories. Some laboratories have been touting an extraction method as a superior type of testing. Not only is this untrue but, in fact, the extraction method has many flaws that may render inaccurate results.

An extraction step is only necessary if the test method employed has a poor sensitivity. For example, if a laboratory uses testing methods designed for serum, which is much less sensitive than the more expensive methods designed for saliva, an attempt to concentrate the hormones is needed in order to measure the hormones. Recall that the serum tests need only measure down to nanogram levels whereas, salivary hormone levels are in the picogram ranges (1000 times less concentrated). On the other hand, if a laboratory uses FDA approved saliva testing methods that have shown the sensitivity down to 1pg/ml, concentrating the sample is not necessary. In fact, adding an extraction step together with using serum test kits leads to many errors and inaccurate results. Labrix testing methods and kits are FDA approved for saliva and show sensitivity to the picogram level.

Extraction methods employed by some salivary hormone testing laboratories introduce errors including cross contamination of samples and dilution issues. Special columns are necessary for extracting samples, and if they are used more than once carryover will cross contaminate the samples that follow. This is a particular problem if a sample with high levels, e.g. a sample contaminated at the time of collection or from someone on topical therapy, is followed by a sample where the provider is looking for baseline levels. Most laboratories use their columns on multiple samples to save money. This should never be done as it leads to serious sample contamination—ask the question. Ask the prospective laboratory how they validated their process. They should have compared their results with other laboratories in a blinded fashion and they should treat any controls, standards or calibrators in exactly the same manner as the patient samples. Be sure to ask for the data to review to your own satisfaction. One of the major drawbacks of saliva testing, and we believe a critical problem, is the lack of validation.

An additional drawback of an extraction method is the large volume of saliva needed for processing. One laboratory requires 7.5cc of sample (10cc tube), which certainly introduces a dilution factor and even more room for error of results—the first saliva out of the mouth has the highest concentration of hormone. The Labrix system requires only 1cc per collection, which only takes 2-3 minutes and we rarely find an individual who cannot provide adequate sample volume. Furthermore, an extraction step limits the number of samples that can be collected for accurate hormone analysis; imagine the inconvenience of trying to collect four 7.5cc samples throughout the day.

The direct testing methods used by Labrix are designed for saliva and have been validated externally by the test kit manufacturer and internally by our technical staff. Our processing methods clear the saliva of contaminants by freezing the sample at time of collection and again once it arrives at the laboratory, which is followed by centrifugation—a crystal clear sample is then directly tested without any adulteration. An equal aliquot from four samples (required from each individual) is pooled to give the average sex hormone level throughout the day. This is critical! Hormone levels vary as often as every ten minutes, so that a single AM sample will at times lead to the wrong conclusion and therefore the inappropriate treatment decision. The cortisol levels, of course, are taken directly from the timed tubes and tested individually.

We highly recommend that if you are considering using a laboratory that employs an extraction step, you do split samples to satisfy yourself with the claims of accuracy. We will gladly supply a split sample protocol.

Following a Sample Through the Lab

In order to fully understand the entire process for saliva testing, let’s follow a specimen from the initial patient appointment to the receipt of laboratory results:

After hearing the symptoms from your patient, you decide to test her salivary hormone levels. You go over the collection instructions with her and mark the appropriate test request on the requisition before sending her home with the kit.

At home, the patient reviews the instructions, fills out the test requisition form, and collects her specimens. She puts everything into the postage paid box and drops it into the mail.

A few days later, the specimens arrive in our specimen processing department. There they are opened and processed one at a time. The specimen processors review each requisition and each specimen for both accuracy and timely collection. Specimens are then submitted and are further processed for testing.

Within one or two business days, the specimens are tested using EIA immunoassay technology. All aspects of the testing are performed by qualified and licensed laboratory personnel. The testing takes from 2-4 hours to complete, depending on the assay. As tests are completed, they are reviewed by the testing personnel under strict quality control rules, and are then passed on to the Laboratory Director for final approval. It is at this point that the test results are available for interpretations by one of our expert physicians. Each test report is reviewed individually and based on the combination of laboratory results, patient symptoms and information provided by the patient, an interpretation is rendered. Interpreters may hold the report and order tests to be repeated in order to confirm clinically inconsistent high or low values. When this occurs, the testing process begins all over again.

The test results are electronically available to the ordering provider as soon as the interpretation is complete. If the provider is not signed up to receive reports online, they will receive their reports the next business day via fax. Once the provider receives their results, they can begin to help their patient balance their hormones.

[Information courtesy of Labrix Clinical Services, Inc. (www.labrix.com)]