Test Overview
Test Methodology


Test Usage

Support the diagnosis of primary atypical pneumonia (Mycoplasma pneumoniae), hemolytic anemia, cirrhosis, Raynaud's disease, some viral diseases, and infectious diseases such as staphylococcemia and influenza. May also occur in pregnancy. A fourfold rising and falling titer is found in primary atypical pneumonia (PAP); 55% of patients with PAP have rising titers. Ninety percent of these are severely affected or have prolonged illness.

Reference Range *

Titer less than 1:40

* Reference ranges may change over time. Please refer to the original patient report when evaluating results.

Test Limitations

False negatives may occur if serum is refrigerated on the clot. Only half of patients with M. pneumoniae infection will have a positive test. Sensitivity: 1:10

Test Details
Days Set Up
Monday, Thursday
Analytic Time

24 hours

Soft Order Code
MiChart Code
Cold Agglutinin Screen
  • Agglutinins, Cold
  • COLD
Chemical Pathology
Specimen Requirements
Collection Instructions

Whole blood must be maintained at 37 degrees C +/- 2 degrees C from the time it is collected until serum is separated from the cells. Collect blood into prewarmed red top tubes (serum separator tube is unacceptable). Within one minute, specimen must be placed into a 37 degrees C +/- 2 degrees C incubator or waterbath. Leave specimen at 37 degrees C until clot retracts; blood may be left at 37 degrees C up to 24 hours. In the laboratory, process as follows: 1) Place specimen in 37 degrees C waterbath or incubator for 30 minutes and allow to clot. 2) After 30 minute incubation at 37 degrees C, centrifuge specimen at 2000 rpm for 5 minutes. 3) Aliquot serum into a plastic vial and refrigerate. 4) Indicate on requisition or manifest that the specimen was incubated at 37 degrees C for 30 minutes. If specimen processing equipment is not available, the patient should have the specimen collected at an Mlabs Blood Drawing station.

Special Handling

Collect specimen in a red top tube; do not use SST tube.
Place specimen in 37 degrees C Thermos containing a warm pack or waterbath immediately. See Onsite Collection Instructions.

Alternate Specimen
Normal Volume
7 mL whole blood
Minimum Volume
3 mL whole blood
Rejection Criteria
Unacceptable specimens include SST tube, specimen that was refrigerated before serum was separated, specimen not allowed to clot at 37C, hemolyzed and chylous serum.
Additional Information

The most common cause of elevated cold agglutinins in high titer is secondary to an infection with Mycoplasma pneumoniae. Cold agglutinins are usually IgM autoantibodies directed against the Ii antigens of human RBCs. These antibodies may be found in patients with cold agglutinin disease or may occur transiently following a number of acute infectious illnesses. Cold agglutinins of cold agglutinin disease are usually monoclonal IgM Kappa. Cold antibodies of IgG, IgA, or IgM type directed against i antigens may be found in infectious mono. Antibodies reacting closer to physiologic temperatures are more likely to be clinically important. In PAP, cold agglutinins are demonstrated one week after onset; the titer increases in 8 to 10 days, peaks at 12 to 25 days, and rapidly falls after day 30. Antibiotic therapy may interfere with antibody formation.

CPT Code
Fee Code