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Real-time
saliva analysis
Why Saliva?
For many years saliva has been used as a biological fluid for the detection of different biomarkers such as electrolytes, hormones, drugs and antibodies in human and veterinary medicine. Indeed, the field of salivary diagnostics has become a billion dollar market in the USA alone, in what is becoming a truly global industry.

Sample collection is non-invasive, painless and very convenient – especially for clients / patients. These are the key advantages to using this mode as opposed to blood, which rapidly
is more cumbersome, invasive, sometimes painful, carries greater risk of infection and in most environments requires medical supervision. Also, for many, the simple notion of blood testing is unsavoury.
Furthermore, collecting saliva samples is possible any time, day or night because of the overall convenience of collection. Saliva collection can be accomplished under circumstances where blood collection is difficult or inadvisable.

Samples can be collected quickly and easily using the SOMA OFC II and analysed within a matter of minutes from collection using the SOMA LFD Reader or Cube Reader and / or sent to the lab where a wider range of tests are available. The SOMA OFC II can be used as a stand alone collection device, even if not using the SOMA Point of Care testing range.

Watch how quick and easy it is for a Professional Footballer to give a sample with the OFC II
COLLECTION SOLUTIONS
The Soma OFC II has improved functionality, making it the best option for saliva collection, whether for real-time, or subsequent analysis. Quick & easy to use, unaffected by recent food and drink; once the swab is in the buffer, your sample is stable for months.
OFC II
Simple Analysis
QUICK & EASY WITH THE CUBE
Watch to see how easy it is to run a test on the SOMA LFD and Cube Reader.
SQUAD TESTING?
Watch the video to see how you can test 30 samples in 25 minutes. With the Cube it is even quicker…!!!
Instant data
SOMA Oral Fluid Collector II (OFC II)
PRODUCT FACTS

SOMA Product Code
SOMA OFC II

Kit Contents
SOMA OFC Swab and SOMA OFC Buffer

Applications

For the collection of saliva samples for analysis either on SOMA LFD Reader or Laboratory analysis. For use in Sport, Exercise, Corporate, Healthcare, Drugs of Abuse Testing, Research and Veterinary environments.

Collection Time
Typically between 20-50 seconds.

Collection Volume
Typically 0.5 mL of oral fluid.

Shelf-Life
OFC Swab 24 Months
OFC Buffer 18 Months

Storage
4˚C to 40˚C

Sample Stability*
At least 2 years at -20˚C
At least 3 months at 4˚C
At least 3 weeks at 37˚C
The SOMA OFC II consists of two parts, a swab for the collection of the sample and a bottle of buffer. The swab can be used for all types of application where collection of saliva / oral fluid is required. Samples can either be analysed immediately on the SOMA LFD, or can be sent to the lab for subsequent processing. The swab consists of specially formulated polymer based materials attached to a small plastic tube which contains a volume adequacy indicator. The indicator shows a clear colour change when 0.5 mL of oral fluid has been collected. It is then placed in the bottle containing the proprietary SOMA buffer. It is recommended that the swab is placed on top of the tongue and the mouth closed while sample collection takes place, so as to recruit oral fluid from all salivary glands. In most individuals, under normal circumstances, collection takes less than a minute to complete.
The SOMA OFC buffer solution comes in different volumes depending upon the application and required sensitivity of the assay in question. The buffer has four main properties to facilitate collection and stability of the samples:
  • 1
    Buffers, to negate the effect of pH variability due to recent food and drink taken.
  • 2
    Preservatives, to prevent growth or micro-organisms, keeping samples stable at room temperature for weeks (longer when refrigerated or frozen).
  • 3
    Extraction agents, to allow immediate recovery of target analytes with less than two minutes of gentle mixing.
  • 4
    Acts as a run buffer for when running tests on the SOMA LFD
Ideal for Point of Care Test
The SOMA OFC II is the perfect partner for the SOMA LFD test range. It is quick and convenient to use in the sporting environment. Mixing of the sample is best done in a rhythmic back and forth, or up and down, motion to facilitate extraction of the target analyte. There is no need for freezing, pipettes, centrifuges or any other laboratory equipment. To run a test on the LFD, simply flip the cap and release two drops from the dropper cap on the bottle.

Laboratory Application

Because samples are very stable in the SOMA Buffer (three weeks at 37˚ C) the OFC is ideal for remote field trials. It has been used for research studies in environmental extremes such as Mount Everest, Siberia, Antarctica and the heat and altitude in Bolivia. This simple noninvasive collection method needs no expert supervision and can simply be posted to a laboratory for subsequent analysis.

Validation
Jehanli et al. (2011) "Development and validation of an Oral Fluid Collection device and its use in the immunoassay of salivary steroids and imunoglobulins in sports persons." Presented at the Inter national Society of Exercise Immunology Symposium 2011.

LFD Readers
Once you have added two drops of sample to your LFD and it has created 2 or 3 red lines, you need to scan it after the incubation time to get a quantitative result.

Test LFDs:
Alpha-Amylase LFD
Cortisol LFD
IgA LFD
IgG LFD
Testosteron LFD





Monitoring Stress
Research shows that there are two distinct systems involved in the stress response of humans and other animals. The response and activity of both systems can be measured via various biomarkers in saliva. The classic method of characterising a stress response is to measure cortisol levels, which rise due to a series of changes in the hypothalamic-piuitary-adrenal (HPA) axis. However, research has shown that this system can be relatively slow in responding to stress (a latency of some 10-15 minutes); whilst changes in the sIgA and alpha-amylase response, markers of the sympathetic nervous system (SNS) activity, are somewhat quicker1. It is also well known that chronic stress with elevated cortisol levels can lead to immune suppression and reduced sIgA values. With repeated measurements within a day the diurnal pattern of all biomarkers can be established quickly and conveniently on the SOMA LFD platform.
The Diurnal Profile

In corporate and health & wellness settings it is more common to look at the diurnal profile of cortisol, typically with 5 samples taken throughout the day.

Typical diurnal profile sample times:

  1. Awakening
  2. Awakening +30 minutes
  3. Midday
  4. Around 5 p.m.
  5. Bedtime
What's "Normal"??

"Normal" values are challenging at the best of times. Even with lab assays, there are a variety of different kits that can be used and they all give different values for the same sample. Added to that, using the same assay kits in different labs will still give variability2.

This is why we provide data using only SOMA kits to help you put your results into context. Independent research has shown SOMA Cortisol tests give similar values to Salimetrics laboratory ELISA kits3.

Cortisol Profiles from 267 individuals (170 male, 97 female) using the SOMA cortisol assay. (Time Points are (1) Awakening; (2) +30 minutes; (3) ~12:00; 4 ~17:00; (5) Bed-time).
Area Under the Curve

The shape and character of the diurnal profile of cortisol can give useful insight. In well functioning individuals there is typically a sharp peak between sample 1 & 2, with minimal values in the evening. The area under the curve (AUC) can be useful to examine the overall stress output. A flat profile implies chronic stress. To measure AUC you need the exact time of day that the samples were taken and the cortisol values using SOMA cortisol assay
Alpha-Amylase

Cortisol is a great biomarker for monitoring chronic stress or catabolic activity in physical contexts, but if measuring acute stress is your thing, then try the protein markers of alpha-amylase or IgA to investigate rapid stress responses. One of the advantages of these tests is although they have a small diurnal pattern; it is not nearly as pronounced as the cortisol diurnal rhythm. This can make these protein markers a good alternative for stress monitoring where time of day (or more appropriately, time since awakening) cannot be so rigidly controlled.

Salivary Alpha-Amylase (sAA) Profiles from 89 individuals (58 male, 31 female) using the SOMA sAA assay. (Time Points are (1) Awakening; (2) +30 minutes; (3) ~12:00; 4 ~17:00; (5) Bed-time).
REFERENCES:

  1. Nomura S (2012) Salivary Hormones, Immunes and Other Secretory Substances as Possible Stress Biomarker, Biomarker, Prof. Tapan Khan (Ed.) p 247-270
  2. Miller R et al., (2013) Comparison of salivary cortisol as measured by different immunoassays and tandem mass spectrometry. Psychoneuroendocrinology, 38: 50-57
  3. Fisher R et al., (2015) The Validity and Reliability for a Salivary Cortisol Point of Care Test. J Athl Enhancement, 4:4
    Monitoring Immune Function
    Soma has developed 3 Point of Care Tests to monitor Immune Function. You can even have 2 tests on the same LFD:

    • IgA
    • IgG
    • Alpha-Amylase
    IgA

    Secretory IgA is a protein, more specifically an antibody which is the first line of defence on the mucosal surfaces against upper respiratory tract and GI tract infection. It is sometimes used as a surrogate marker of the general immune system, although it should be remembered that the immune system is multi-faceted and somewhat complex in nature.

    Research in athletes has shown that if sIgA levels drop significantly from an individual's baseline norm then they have a one in two chance of an infection in the following two to three weeks (Neville et al., 2008; Dunbar et al., 2013). This is a hard-hitting fact.

    Outside of sport, the use of a simple immune marker can be used to deal with growing problems of absenteeism from and presenteeism at work. The latter is where people go to work in ill-health and productivity is low not just on those days, but for many days after as a result. In the military, rapid insight into an individual's immune status can be advantageous in assessing response to various exercises or combat, as well as examining risk of future illness.
    IgG

    Salivary IgG is another antibody, which is not usually secreted in salivary glands, but diffuses from the blood. In fact IgG is the main immunoglobulin in human blood. However you can measure the concentration in saliva using the Soma IgG LFD test. It is not commonly used as an isolated marker, but has been assessed in ratio form with sIgA (Dunbar et al., 2013) and improved the monitoring of illness as opposed to using just IgA alone. Along with IgM this is the other key immunoglobulin found in saliva, but in contrast to sIgA has been investigated in very few research studies. One study has found that salivary IgG levels remain unchanged during acute exercise in both athletic and healthy participants.
    Alpha-Amylase

    Salivary Alpha-amylase (sAA) is another antimicrobial protein in saliva. It is more commonly known as a key digestive enzyme responsible for breaking down starches in the mouth and gut. However, its role in protection against disease is its ability to inhibit growth and adherence of bacteria to epithelial surfaces.

    sAA has appeared increasingly in the literature as an acute stress biomarker, reflecting sympathetic nervous system activity and differs from cortisol which is slower to respond being a function of HPA-activity. Thus the analyte has been used frequently as a rapid acting acute stress marker in the behavioural sciences. Like cortisol, it is very much viewed as a biomarker reflecting the stress response to exertion, yet some argue that sAA is a more sensitive marker, partly because it is quicker to spike after stressful activity, than the more slowly acting cortisol.
    Monitoring Readiness to Train
    There are several Soma Biomarkers that you can use to monitor Readiness to Train:

    • IgA
    • Alpha-Amylase
    • Cortisol

    Always remember that "norms" don't exist for these salivary biomarkers; it is all about identifying individual norms and looking for deviations from these.
    IgA

    When the body comes under pressure from a variety of stressors on a chronic basis, the immune system can become suppressed. It is at this point that the body is more vulnerable to pathogens to which the individual is exposed. In sport, one typical cause of this could be consistently high training loads, where an individual is not fully recovered. In individual sports, there is a strong argument for regular monitoring of IgA, so close tabs can be kept on how that individual is recovering. This monitoring could be daily or even twice daily (before and after practice) because research shows that although moderate exercise can be an immune booster, intense exercise can have the opposite effect i.e. suppress the immune system. Tracking responses to various workouts or competitions can be useful in gaining an insight in to how an athlete copes or recovers. Other stressor can also have an effect on IgA, such as travel and lifestyle factors such as poor sleep, stress (of an emotional or psychological nature) and in the student athlete, academic load. Significant changes in any of these factors will see changes in the baseline norm established for each individual.

    Individual profiles

    The development of individual profile is essential for the interpretation of data on all SOMA tests. It is usual that a minimum of 5 to 6 tests are required to establish a baseline for any individual athlete. That baseline will of course grow with subsequent testing and the greater your data set the more robust your norms become.
    Research has shown that if sIgA levels drop significantly from an individual's baseline norm then athletes have a one in two chance of an infection in the following two to three weeks (Neville et al., 2008; Dunbar et al., 2013). This is a hard-hitting fact.
    In team sports, measurement of sIgA is not required so frequently. A perfectly good monitoring system would test players once or twice a week and again referencing individual norms and deviations from such, to contribute in creating a "readiness to train" index.

    The chart shows weekly IgA values plotted with the mean (solid line) and 40% above and below (dashed line). Outliers are easy to identify.
    Peaks and Troughs

    During monitoring it can be seen that on occasion, IgA levels can spike from individual normal levels. This will be the result of one of two factors. Either the individual is fighting an infection (remember that they can be fighting an infection but may not be symptomatic, for example if a relative or housemate has an infection) or when they become acutely stressed. It is quite common in some countries such as Japan, that sIgA is used as an acute stress marker. This can be seen in studies investigating exam stress. We have also seen spikes in IgA in some (but not all) individuals before important competition.


    Analysing a Squad

    Once you have developed norms for each athlete you can see readiness to train easily in a squad. In the chart below, each black column represents an athlete's normal range. The red marks are the values for that day, so it is quick and easy to see who is out of range in a squad.
    Alpha-Amylase

    Salivary alpha-amylase (sAA) has appeared increasingly in the literature as an acute stress biomarker, reflecting sympathetic nervous system activity and differs from cortisol which is slower to respond being a function of HPA-activity. Thus the analyte has been used frequently as a rapid acting acute stress marker in the behavioural sciences. Like cortisol, it is very much viewed as a biomarker reflecting the stress response to exertion, yet some argue that sAA is a more sensitive marker, partly because it is quicker to spike after stressful activity, than the more slowly acting cortisol. sAA has been shown to be an alternative marker to determine anaerobic threshold, but the cost of such measurement, as opposed to the cheaper blood lactate, would leave it as a questionable, option where repeated measurements are taken within the test. However, it does have several possibilities as an acute physiological marker in the sport and exercise environment.
    There is a strong argument for the use of sAA response to monitor adaptations to training on a longitudinal basis, because levels are likely to be suppressed in elite athletes during heavy training periods. Logic dictates that the hard training periods are where sympathetic withdrawal of the nervous system is likely to happen during non-functional over-reaching. Research is still in its infancy with regards to such monitoring, but certainly gathering momentum and is the focus of much work at Bond University in Gold Coast, Australia. It could certainly be viewed as the "new kid on the block" as regards stress biomarkers. As such, other uses could be for monitoring stress in military and other stressful environments, such as fire fighters, police forces etc.
    We have already demonstrated sAA is useful for the construction of an IgA / Amylase ratio which shows better capability for predicting URTI in a cohort of Premier League academy soccer players (presented at International Society of Exercise Immunology Symposium, Newcastle, NSW, Australia in 2013). The IgA / sAA ratio is becoming a very popular biomarking tool in elite soccer teams in the UK, an elsewhere in Europe, as well as a wide range of sports in Australia.
    Cortisol

    Cortisol, being a catabolic hormone, is commonly measured in sporting environments when trying to categorise responses to different types of training. The theory being that if cortisol is high, then the body is in a catabolic state and thus readiness to train is impaired. Timing of sampling needs to be controlled and factored into the assessment of data due to the strong circadian pattern. Cortisol is often evaluated in context with an anabolic hormone to give a ratio. In sport the hormone of choice is typically Testosterone and in corporate worlds DHEA is used. There is no clear reason for the choice of each anabolic hormone, after all DHEA is a precursor to testosterone. It is well known that testosterone does tend, in general, to be significantly lower in females than males and the DHEA levels tend to be lower with increasing age.
    In sport situations where cortisol is monitored routinely, higher levels would tend to indicate greater catabolic breakdown of muscle with acute fatigue. However, during chronic fatigue there may be a situation where cortisol levels tend to be much lower than normal and thus represent a suppressed state.

    In team sports, the squad will usually be tested once or twice per week, in an attempt to monitor recovery status from competitive games or during intense training periods. With individuals, the testing may become a little more frequent and can even be on a daily basis. It is important to remember that, being a stress hormone, both psychological / emotional factors will affect cortisol values as well as physical factors. Thus there is a requirement of an interdisciplinary approach to data interpretation. Like IgA and other analytes there is no clear reference value to aim for, it is a question of again establishing a baseline and examining relative changes form the baseline established for each individual.

    This data below shows a strong correlation between group mean salivary cortisol (using the IgA / Cortisol Dual LFD) and group mean HML Distance (Training Load) through half a Premier League season in soccer players.