- Red top tube
- 7 mL of venous blood
Additional information
- Keep the sample warm
- Do not refrigerate
- Cold agglutinins are autoantibodies, which are capable of agglutinating red blood cells (RBCs) at temperatures below 35°C
- These antibodies can be monoclonal or polyclonal, and have been associated with various infections and neoplasms, most commonly with atypical pneumonia caused by Mycoplasma pneumoniae
- The diagnosis depends on demonstrating a 4-fold or higher increase in antibody titers between an early acute-phase blood serum sample and a blood serum sample taken in the convalescence phase, 7–10 days after the first sample
- Cold agglutinins may be detected in asymptomatic individuals during routine blood typing and crossmatching. In these individuals, there is a positive indirect antiglobulin (Coomb's) test, which detects free serum antibodies against RBCs
- The cold agglutinin test is done to:
- Aid in the early detection and confirmation of Mycoplasma pneumoniae infection
- Determine whether high titer levels of cold agglutinins are causing autoimmune hemolytic anemia which may be due to:
- Viral infections
- Cancer (lymphoma, multiple myeloma)
- Rheumatoid arthritis
- Scleroderma
- Systemic lupus erythematotosus
- Idiopathic cold agglutinin disease
- Assist in the confirmation of
- Influenza
- Atypical pneumonia caused by Mycoplasma pneumoniae
- Pulmonary embolus
- Evaluate symptoms on exposure to low temperature such as numbness, burning pain, pale skin of the fingertips, toes, ears, or nose
- When rouleaux formation is noted on complete blood count test
- Patients with cold agglutinins may present with vascular abnormalities including:
- Mottled skin
- Purpura
- Jaundice
- Pallor
- Pain or swelling of extremities
- Cramping of fingers and toes
- Hemoglobinuria
- The rise in titer during the course of illness is more important than any single high value. The titer usually decreases by 4–6 weeks after the onset of illness
- Patients with cold agglutinins develop 2 types of antibodies, anti-I and anti-i, which are usually of IgM class, but may be of IgG, IgA, or mixed IgM-IgG classes. These antibodies react with the I or i antigens on adult human type O red cells at temperatures below 35°C, resulting in agglutination
- Cold agglutinins develop in approximately 34-68% of patients with M. peumoniae infection. M. pneumoniae has I antigen specificity to human RBC membranes to which the antibodies develop. These antibodies arise before the complement fixing antibodies, and are useful in early detection
- The rapid cold agglutination test has positive predictive value (PPV) of 70% in mycoplasma infection. Thus, in children and young adults with clinical signs of M. pneumoniae infection and positive cold agglutination test, a presumptive diagnosis can be made
- In mycoplasma pneumonia, the titer rises 8–10 days after onset, peaks in 12–25 days, and decreases 30 days after onset. Up to 90% of people with severe illness exhibit positive titers
- The presence of cold agglutinins in the blood can interfere with automated machines that measure blood count
- Blood that contains high levels of cold agglutinins may be difficult to Type properly
- Recent use of antibiotics may interfere with cold agglutinin development
- Cephalosporins
- Penicillins
Additional information
- If cold agglutinin disease is suspected, keep patient warm. If patient is exposed to low temperatures, agglutination may occur within peripheral vessels leading to frostbite, anemia, Raynaud's phenomenon, and focal gangrene.
- False low titer is seen in samples which have been refrigerated, as cold agglutinins will coat RBCs leaving none in the serum
- Related laboratory tests include:
- Arterial/alveolar oxygen ratio
- Blood gases
- Complete blood count
Consult your laboratory for their normal ranges as these may vary somewhat from the ones listed below.
- Qualitative: Negative
- Quantitative: <1:32
- Serial Quantitative: Less than a four fold increase in titer in serial blood draws over 7-10 days
- Extremely high titers (e.g. >1:2,000) are seen in idiopathic cold agglutinin disease that precedes lymphoma development
- Conditions which are associated with elevated cold agglutinins titers, include:
- Polyclonal
- Mycoplasma pneumoniae infection
- Collagen vascular diseases
- Cytomegalovirus infection
- Infectious mononucleosis (Epstein Barr Virus)
- Listeriosis
- Mumps
- Subacute bacterial endocarditis
- Syphilis
- Systemic lupus erythematosus
- Tropical diseases
- Malaria
- Tropical Eosinophilia
- Trypanosomiasis
- Monoclonal
- Cold agglutinin disease
- Waldenström's macroglobulinemia
- Hodgkin's lymphoma
- Non-hodgkin's lymphoma
- Chronic lymphocytic leukemia
- Chronic myelogenous leukemia
- Cold antibody hemolytic anemia
- Kaposi's sarcoma
- Multiple myeloma
- Paroxysmal cold hemoglobinuria
- Plasmacytoma
- Severe combined immunodeficiency with B-lymphocyte proliferation
- Severe Raynaud's phenomenon (may lead to gangrene)
- Transient increase in titers are associated with
- Cytomegalovirus (CMV) infection
- Epstein Barr Virus (EBV) infection [Infectious mononucleosis)
- Hepatic cirrhosis
- Measles
- Mumps
- Syphilis (Congenital)
- Viral pneumonia
- Trypanosomiasis
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- ChangYT et al. The significance of a rapid cold hemagglutination test for detecting mycoplasma infections in children with asthma exacerbation. J Microbiol Immunol Infect. 2006 Feb; 39(1):28-32.
- Ciejka JZ et al. Severe cold agglutinin disease and cryoglobulinemia secondary to a monoclonal anti-Pr2 IgM lambda cryoagglutinin. Clin Exp Rheumatol. 1999 Mar-Apr;17(2):227-31
- De Boer J et al. Severe diffuse interstitial pneumonia due to Mycoplasma pneumoniae in a patient with respiratory insufficiency. Neth J Med. 2003 Mar;61(3):91-4
- GehrsBC, et al. Autoimmune hemolytic anemia. Am J Hematol. 2002 Apr;69(4):258-71
- MedlinePlus Medical Encyclopedia®. Febrile/cold agglutinins.[Homepage on the Internet]©2005. Last updated on April 20, 2005. Last accessed on October 20, 2006. Available at URL: http://www.nlm.nih.gov/medlineplus/ency/article/003549.htm
- Navid Madershahian et al. Cold Agglutinins In On-Pump Cardiac Surgery: A Rare But Potentially Lethal Problem. The Internet Journal of Perfusionists. 2004. Volume 2 Number 1.
- Oshima M et al. Low-titer cold agglutinin disease with systemic sclerosis. Intern Med. 2004 Feb;43(2):139-42
- Stone MJ et al. Autoantibody activity in Waldenstrom's macroglobulinemia. Clin Lymphoma. 2005 Mar;5(4):225-9
- Teachey DT et al. Development of cold agglutinin autoimmune hemolytic anemia during treatment for pediatric acute lymphoblastic leukemia. J Pediatr Hematol Oncol. 2005 Jul;27(7):397-9
- Torres JR, et al. Low-grade parasitaemias and cold agglutinins in patients with hyper-reactive malarious splenomegaly and acute haemolysis. Ann Trop Med Parasitol. 2003 Mar;97(2):125-30