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Equine SAA handbook

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Equine Serum Amyloid A (SAA) picks up on a systemic inflammation when other signs may be unclear.

Use Serum Amyloid A

A high Serum Amyloid A value should be an indication to start treatment for inflammation relapse, even if other signs are absent.

4 To determine the extensiveness of the inflammation.

4 To determine if selected treatment is effective.

4 As a rule in/rule out marker for inflammatory diseases.

4 To monitor the post-operative effects and recovery after surgery.

Near Patient Testing has been an increasing trend within human medicine the last years. Veterinary medicine is starting to embrace this as well.

The benefits of having a system that simplifies diagnosis, monitors treatment and is specifically designed to do so right next to the patient in “real” time are huge.

For an efficient work-flow in an emergency setting, the test must have a short turn around time, optimally less than 15 minutes. Additionally, the test-format has to be easy-to-use so that non-laboratory personnel are able to perform the test.

The immediate detection of an inflammatory response and the monitoring of it’s clinical course are primary challenges for veterinary medicine.

Systemic inflammation can be initiated by bacteria, viruses, parasites, fungi, neoplasia and trauma.

The search for early inflammation markers has therefore been an important focus in veterinary medical research. Special attention has been on the identification of biochemical parameters that have the sensitivity and specificity to both signal the presence and evaluate the intensity of an inflammatory response.

Acute Phase Proteins are a group of biological markers that have a direct response to a systemic inflammation and a fast decrease in concentration upon removal.

Serum Amyloid A belongs to the group of major acute phase proteins in horses. I.e. the serum concentration of SAA increases more, as a result of systemic inflammation, compared to other acute phase proteins in horses (Ref. 1,2,3).

Acute Phase Proteins in Veterinary Practice

Easy to use point-of-care testing

LifeAssays® Equine SAA System LifeAssays® Equine SAA System

4 Specific and objective marker for systemic inflammation.

4 Real time marker – starts after 6h–12h, peaks after 36h (Ref. 4,5).

4 Large diagnostic window – multifold increase in concentration as compared to reference range (Ref. 3).

4 An elevated serum value always indicates pathology.

4 Use SAA to monitor the post-operative effects and recovery after surgery (Ref. 6).

Conditions and diseases stimulating a systemic inflammation may be monitored, for example: Bacterial infections and viral infections (Ref. 1,10)

Key benefits of Serum Amyloid A (SAA) as “the Systemic Inflammatory Marker” in horses

LifeAssays® Equine SAA System

Soft tissue Surgery Orthopedic Surgery

TimeTime

CRP CRP

Surgical procedure Surgical procedureHours

SAA

Admission Pre-diagnostic value

Post-diagnostic value

Administrate treatment

To best make use of a post-diagnostic SAA value it is of great importance to have a pre-diagnostic value, since this will enable you to compare the two.

0 10


Performance data

Linear range 10 – 3000 mg/L

Imprecision CV= 7%

Correlation r2 = 0.95 vs Eiken Serum Amyloid A

Cut-off (normal/abnormal)

20 mg/L

LifeAssays® Equine SAA system consists of a bench-top instrument (LifeAssays® Magnia Reader), single-use reagent tests and a disposable calibration keycard. All reagent identification data, as well as, a self-executable algorithm is contained on the disposable calibration keycard providing better traceability and improved system control.

The keycard is inserted into the LifeAssays® Magnia Reader when performing a test. Thus, reagent/software upgrades are easily provided with each new reagent kit.

LifeAssays® Equine SAA System

LifeAssays® Equine SAA test kit provides a point-of-care quantitative measurement of Equine SAA in serum, in 5 minutes. This immunochromatographic assay uses superpara magnetically labeled antibodies for detection.

The LifeAssays® Equine SAA test kit helps veterinary professionals to easily integrate SAA measurements as a point-of-care diagnostic tool to diagnose and monitor disease progression as well as treatment efficiency on inflammations in horses.

Repeating a LifeAssays® Equine SAA test during and after treatment (e.g. antibiotics) will show if the selected treatment has been effective and reduced the inflammation or infection, see next page.

Repeating a LifeAssays® Equine SAA test after surgery will show recovery and ascertain that no post- operative inflammation is at hand.

1. Pepys MB, Baltz ML, Tennent GA, Kent J, Ousey J, Rossdale PD. Serum amyloid A protein (SAA) in horses: objective measurement of the acute phase response. Equine Vet J. 1989;21(2):106–109.

2. Hulten C, Tulamo RM, Suominen MM, Burvall K, Marhaug G, Forsberg M. A non-competitive chemiluminescence enzyme immunoassay for the equine acute phase protein serum amyloid A (SAA) - a clinically useful inflammatory marker in the horse. Vet Immunol Immunopathol. 1999;68(2-4):267–281. doi: 10.1016/S0165-2427(99)00027-6.

3. Cray C, Zaias J, Altman NHet al Comp Med. 59(6): 517–526, 2009 Dec ;. Acute Phase Response in Animals: A Review.

4. Jacobsen S. AAEP Proceedings, vol 53, 2007 Review of Equine Acute-Phase-Proteins.https://aaep.org/sites/default/files/issues/proceedings-07proceedings-z9100107000230.pdf

5. Tape C, Kisilevsky R. Apolipoprotein A-I and apolipoprotein SAA half-lives during acute inflammation and amyloidogenesis. Biochim Biophys Acta. 1990;1043(3):295–300.

6. Jacobsen S, Nielsen JV, Kjelgaard-Hansen M, Toelboell T, Fjeldborg J, Halling-Thomsen M, Martinussen T, Thoefner MB. Acute phase response to surgery of varying intensity in horses: a preliminary study. Vet Surg. 2009;38(6):762–769. doi: 10.1111/j.1532-950X.2009.00564.x

7. Hobo S, Niwa H, Anzai T. Evaluation of serum amyloid A and surfactant protein D in sera for identification of the clinical condition of horses with bacterial pneumonia. J Vet Med Sci. 2007;69(8):827–830. doi: 10.1292/jvms.69.827

8. Jacobsen S, Thomsen MH, Nanni S. Concentrations of serum amyloid A in serum and synovial fluid from healthy horses and horses with joint disease. Am J Vet Res. 2006;67(10):1738–1742. doi: 10.2460/ajvr.67.10.1738.

9. Paltrinieri S, Giordano A, Villani M, Manfrin M, Panzani S, Veronesi MC. Influence of age and foaling on plasma protein electrophoresis and serum amyloid A and their possible role as markers of equine neonatal septicaemia. Vet J. 2008;176(3):393–396. doi: 10.1016/j.tvjl.2007.05.018.

10. Hulten C, Sandgren B, Skioldebrand E, Klingeborn B, Marhaug G, Forsberg M. The acute phase protein serum amyloid A (SAA) as an inflammatory marker in equine influenza virus infection. Acta Vet Scand. 1999;40(4):323–333.

LifeAssays® Magnia ReaderTechnical Data:

• Easy-to-read display• Bench-top instrument• Calibration keycard• Automatic measurement and

result presentation• Superior non optical system• Battery powered

LifeAssays® Equine SAA Technical Data:

• Easy-to-use• Quantitative: clear relation of

results vs. cut-off: 20 mg/L• Accurate• Rapid : assay time 5 minutes• Small sample volume: 3 µL • Linear range 10–3000 mg/L


References

Short Guide for Easy and Quick Turn-Around Time


Step 1. Turn on the Magnia® Reader. Insert the lot-specific LifeAssays® Equine SAA “calibration key” card into the Reader label pointing up. The Reader then displays “eSAA”, the LOT number and Expiration date.

Step 2. Open the aluminium test pouch and insert the LifeAssays® test strip into the Reader (small arrow on the end of the test device is visible and pointing towards you). The sample well marked with “S” will remain outside the Reader.

Insert test device

Press Power

2Insert Card

1

Press Measure

x30

Insert test device

Press Power

2Insert Card

1

Press Measure

x30

Step 3. Collect the serum/plasma sample using the 3 µl glass capillary and fill completely. Wipe off excess fluid with a clean tissue. Place the capillary into a buffer vial and mix by inverting the tube at least 30 times.

Insert test device

Press Power

2Insert Card

1

Press Measure

x30

Step 4. Add 3 drops of the diluted sample to the sample well marked with “S” with the pipette and immediately press “MEASURE”. The instrument will incubate the LifeAssays® test device for 5 minutes. After 5 minutes the Reader will read and calculate the result. If the Reader is unused for over 20 min, it will turn itself off. The last measured result is not lost but will be displayed when the Reader is powered up again.

Insert test device

Press Power

2Insert Card

1

Press Measure

x30

Step 5. The final result reports as mg/L and prompts: “Remove Test”. When a new LifeAssays® test device is inserted into the Reader the test result will disappear and be permanently lost.

Insert test device

Press Power

2Insert Card

1

Press Measure

x30

Step 6. After recording the result, remove the LifeAssays® test device from the Reader and check the control line visually. A pink control line in window area next to the letter “C” appears. If there is no control line, the test result is invalid and should be discarded.

Insert test device

Press Power

2Insert Card

1

Press Measure

x30

Short Guide for MagniaReader Set-up


Before the Reader can be taken into use after storage or transportation, its battery must be connected and recharged (for recharging see chapter 5.3 in the MagniaReader Operating manual). When you take the reader in use for the first time you have to connect the battery.

To connect the battery, 1. Turn the Reader upside down. The battery hatch is in the bottom of the Reader. 2. Remove the screw of the battery hatch. 3. Turn the Reader over to make the battery come loose. 4. Connect the battery connector. 5. Lower the battery in its place. 6. Close the battery hatch with the screw.

Connecting Battery

To start up the Reader, 1. Push the Power button on the Reader panel. The Reader starts up. 2. Check the display for warning of low battery. If the battery is low, recharge the

battery as instructed in chapter Recharging battery. 3. Remove the piece of cardboard protecting the swingbed. NOTE: Do not throw

the piece of cardboard away. Save it for later use when the Reader is being transported or stored.

4. Make sure the swing bed is released to downward position so that the carrier will not damage the sensor head.

5. Make sure you are holding the carrier the right way. The thicker head of the carrier goes in the Reader first.

6. Push the carrier gently into the Reader until it locks in place. Push the carrier in a slanted position so that the head that goes in first is slightly higher than the other end of the carrier.

Installing carrier

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