Ham’s (acid hemolysin) test

Alternative names 
Ham test

The Ham’s (acid hemolysin) test looks for increased fragility of red blood cells in mild acid.

How the test is performed

Blood is drawn from a vein, usually from the inside of the elbow or the back of the hand. The puncture site is cleaned with antiseptic, and an elastic band or blood pressure cuff is placed around the upper arm to restrict blood flow through the vein. This causes vein to swell with blood.

A needle is inserted into the vein, and the blood is collected in an air-tight vial or a syringe. During the procedure, the band is removed to restore circulation. Once the blood has been collected, the needle is removed, and the puncture site is covered to stop any bleeding.

In infants or young children:
The area is cleansed with antiseptic and punctured with a sharp needle or a lancet. The blood may be collected in a pipette (small glass tube), on a slide, onto a test strip, or into a small container. Cotton or a bandage may be applied to the puncture site if there is any continued bleeding.

How to prepare for the test

There is no special preparation needed for this test.

How the test will feel
When the needle is inserted to draw blood, some people feel moderate pain, while others feel only a prick or stinging sensation. Afterward, there may be some throbbing.

Why the test is performed

This test is used to confirm the diagnosis of paroxysmal nocturnal hemoglobinuria (PNH).

Diagnosis of PNH can be confirmed by having a positive acidified serum test (Ham test). In acidified serum, complement is activated by the alternate pathway. It binds to red blood cells, and ruptures the abnormal PNH cells, which are unusually susceptible to complement. With newer methods of diagnosis, such as flow cytometry, this test has become less important in the diagnosis of PNH.

The Ham test is also positive in another rare disorder called congenital dyserythropoietic anemia, but in this case the sugar-water fragility test is negative. Furthermore, the clinical aspects of this disorder are not similar to PNH.

Normal Values
A negative test is normal.

What abnormal results mean

  • Paroxysmal nocturnal hemoglobinuria  
  • Congenital dyserythropoietic anemia (anemia associated with a hereditary problem with erythropoietin, the hormone that normally triggers red blood cell production)

What the risks are

  • Excessive bleeding  
  • Fainting or feeling light-headed  
  • Hematoma (blood accumulating under the skin)  
  • Infection (a slight risk any time the skin is broken)  
  • Multiple punctures to locate veins

Special considerations
Veins and arteries vary in size from one patient to another, and from one side of the body to the other. Obtaining a blood sample from some people may be more difficult than from others.

Johns Hopkins patient information

Last revised: December 8, 2012
by Brenda A. Kuper, M.D.

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