hemoglobin F in blood (Kleihauer-Betke test)

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When blood smears are immersed in acid buffer, adult hemoglobin is eluted from the erythrocytes, whereas fetal hemoglobin is not. If blood smears are treated in this manner & subsequently stained, erythrocytes having hemoglobin F will take up the stain, while those containing only adult hemoglobin appear as 'ghosts'.

Clinical significance

Fetal hemoglobin estimations are sometimes made to determine possible hemorrhage in the newborn, particularly in cases where there are signs of rectal bleeding. Hemoglobin F assay is also applied to adult as an aid in diagnosing certain types of anemia. For example,10-90% fetal hemoglobin is encountered in patients with thalassemia major. Moreover, small increases of fetal blood pigment is usually observed in patients with sickle cell disease.

It is becoming increasingly common in cases of Rh incompatibility to suppress immune reactions to red blood cells entering maternal circulation from the fetus. The amount of specific gamma globulin, containing anti Rh[D] to be administered, is calculated by assessing the magnitude of fetal-maternal hemorrhage.

Distribution of hemoglobin F within red cells. HbF shows 3 distribution patterns: both HbA & HbF are present in strictly separated cell populations; both hemoglobins are present in equal concentrations in all cells; & both hemoglobins are irregularly distributed within the cells, some cells containing only HbA, some only HbF, & some varying amounts of both.

Below are distribution patterns of HbF & HbA in red cells.

  • Two separate cell populations, HbF containing cells & HbA containing cells.
  • Even distribution of HbA & HbF within cells.
  • Irregular distribution of HbA & HbF within cells, some cells containing more HbF than others & some containing none.



Patient preparation: No special patient preparations required. Fresh capillary or oxalated blood, a minimum of 1 mL. Specimens allowed to stand overnight may give false positive results.

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  1. Package Insert, Fetal Hemoglobin, Sigma Diagnostics, St. Louis, M0, 1988.
  2. Tietz, Norbort W.: Textbook of Clinical Chemistry, W.B. Saunders Company, Philadelphia, PA, p. 1555, 1986.
  3. Bauer, John D. M.D.: Clinical Laboratory Methods, The C.U Mosby Company, St. Louis, MO, pp. 108-109, 1974.
  4. Oski, F.A., Naiman, J.L.: Histologic Problems in the Newborn, 2nd Ed. W.B. Saunders Company, Philadelphia, PA, pp. 62-63, 1972.
  5. Fetal Hemoglobin, APT Test Laboratory Test Directory ARUP: http://www.aruplab.com/guides/ug/tests/0020156.jsp
  6. Kleihauer-Betke Stain Laboratory Test Directory ARUP: http://www.aruplab.com/guides/ug/tests/0040105.jsp
  7. Panel of 10 tests Laboratory Test Directory ARUP: http://www.aruplab.com/guides/ug/tests/0050610.jsp
  8. Panel of 3 tests Laboratory Test Directory ARUP: http://www.aruplab.com/guides/ug/tests/0050613.jsp
  9. Hemoglobin F Laboratory Test Directory ARUP: http://www.aruplab.com/guides/ug/tests/0081348.jsp
  10. Panel of 10 tests Laboratory Test Directory ARUP: http://www.aruplab.com/guides/ug/tests/0000000.jsp