Lab Med Online.  2014 Jul;4(3):152-156. 10.3343/lmo.2014.4.3.152.

Alterations of Complement C3 and C4 Levels in Delayed Testing

• Lee ZY
• Jearn LH
• Park IK
• Kim TY

• KMID: 1706852
• DOI: http://doi.org/10.3343/lmo.2014.4.3.152

Abstract

BACKGROUND
In vitro levels of complement C3 and C4 proteins are sensitive to storage conditions. To avoid in vitro complement activation when testing is delayed, serum should be frozen at -20degrees C within 2 hr of venipuncture. However, this is impractical in routine laboratory work. Therefore, we investigated alterations in C3 and C4 levels in refrigerated specimens over time and derived formulae to estimate initial levels of complement concentrations in delayed testing.
METHODS
Ten fresh specimens were measured for C3 and C4 concentrations and were refrigerated at 4degrees C. We measured C3 and C4 levels in refrigerated samples daily for 4 days using an automated nephelometer (Beckman Coulter Inc., USA).
RESULTS
C3 and C4 levels were significantly increased over time in refrigerated specimens (P<0.001, P<0.001, respectively). The increments in C3 and C4 levels were described by the equations: C3 (mg/dL)=3.55x+87.18 (r=0.9909), and C4 (mg/dL)=0.72x+22.3 (r=0.9395), where x=the number of days samples were refrigerated before testing. Increases in C3 and C4 concentrations were described on a percentage basis by the equations: DeltaC3 (%)=4.14x+1.07 (r=0.9903), and DeltaC4 (%)=3.57x+2.48 (r=0.9405).
CONCLUSIONS
As the measured C3 and C4 concentrations increased by 3.55 mg/dL (4.1%) and 0.72 mg/dL (3.6%) per day in refrigerated specimens, the levels of C3 and C4 should be adjusted in delayed testing. We proposed that the formulae presented be used to back-calculate initial levels of C3 and C4 concentrations.

Keyword

Complement C3; Complement C4; Delayed testing

MeSH Terms

Complement Activation
Complement C3*
Complement C4
Complement System Proteins
Phlebotomy
Complement C3
Complement C4
Complement System Proteins

Figure

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  Fig. 1 Alterations of complement C3 (A) and complement C4 (B) levels in refrigerated specimens over time, expressed as mg/dL.

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  Fig. 2 Alterations of complement C3 (A) and complement C4 (B) levels in refrigerated specimens over time, expressed as % changes.

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