Factors Known to Cause False Positive HIV Antibody Test Results

By Christine Johnson

Continuum Sept./Oct. 1996

The AIDS establishment has managed to convince many people that the HIV antibody tests (ELISA, IFA and Western blot) are "99.5% accurate". In this article Christine Johnson from HEAL Los Angeles, lists conditions documented in the scientific literature known to cause positives on these tests, and gives her references.

It is expected that this list will generate much discussion and dissension. For the time being, a few clarifications should be made at the outset.

Just because something is on the list doesn't mean that it will definitely, or even probably, cause a false-positive. It depends on what antibodies the individual carries; as well as the characteristics of each particular test kit.

For instance, some, but not all, people who have had blood transfusions, prior pregnancies or an organ transplant will make HLA antibodies. And some, but not all, test kits (both ELISA and Western blot) will be contaminated with HLA antigens to which these antibodies can react. Only if these two conditions coincide might you get a false-positive due to HLA cross-reactivity.

Some things are more likely than others to cause false-positives. And some things that we aren't aware of yet, but which may be documented in the future, may cause false-positives. Some of the factors on the list have been documented only for ELISA, some for both ELISA and Western blot (WB).

Some people may be eager to argue that if a factor is only known to cause false-positives on ELISA, this problem won't be carried over to the WB, so everything should be OK. But remember, a WB is positive by virtue of accumulating enough individual positive bands to add up to the total required by whatever criteria you use to interpret it (39) So the more exposures a person has had to foreign antigens, proteins and infectious agents, the more various antibodies he or she will have in their system, and the more likely it is that there will be several cross-reacting antibodies, enough to make the WB positive.

It is to be noted that all AIDS risk groups (and Africans as well), but not the general US or Western European population, have this problem in common: they have been exposed to a plethora of foreign antigens and proteins. This is why people in the AIDS "risk groups" tend to have positive WBs (i.e., to be considered "HIV-infected") and people in the general population don't. However, even people in the low-risk populations may have false-positive Western blots for poorly understood reasons.(47)

Since false-positives to every single HIV protein have been documented (36), how do you know the positive WB bands represent the various proteins to HIV, or just a collection of false-positive bands reacting to several different non-HIV antibodies?

Factors Known to Cause False-Positive HIV Antibody Test Results

  • Anti-carbohydrate antibodies (52, 19, 13)
  • Naturally-occurring antibodies (5, 19)
  • Passive immunization: receipt of gamma globulin or immune globulin (as prophylaxis against infection which contains antibodies)(18, 26, 60, 4, 22, 42, 43, 13)
  • Leprosy (2, 25)
  • Tuberculosis (25)
  • Mycobacterium avium (25)
  • Systemic lupus erythematosus (15, 23)
  • Renal (kidney) failure (48, 23, 13)
  • Hemodialysis/renal failure (56, 16, 41, 10, 49)
  • Alpha interferon therapy in hemodialysis patients (54)
  • Flu (36)
  • Flu vaccination (30, 11, 3, 20, 13, 43)
  • Herpes simplex I (27)
  • Herpes simplex II (11)
  • Upper respiratory tract infection (cold or flu)(11)
  • Recent viral infection or exposure to viral vaccines (11)
  • Pregnancy in multiparous women (58, 53, 13, 43, 36)
  • Malaria (6, 12)
  • High levels of circulating immune complexes (6, 33)
  • Hypergammaglobulinemia (high levels of antibodies) (40, 33)
  • False positives on other tests, including RPR (rapid plasma reagent) test for syphilis (17, 48, 33, 10, 49)
  • Rheumatoid arthritis (36)
  • Hepatitis B vaccination (28, 21, 40, 43)
  • Tetanus vaccination (40)
  • Organ transplantation (1, 36)
  • Renal transplantation (35, 9, 48, 13, 56)
  • Anti-lymphocyte antibodies (56, 31)
  • Anti-collagen antibodies (found in gay men, haemophiliacs, Africans of both sexes and people with leprosy)(31)
  • Serum-positive for rheumatoid factor, antinuclear antibody (both found in rheumatoid arthritis and other autoantibodies)(14, 62, 53)
  • Autoimmune diseases (44, 29, 10, 40, 49, 43): Systemic lupus erythematosus, scleroderma, connective tissue disease, dermatomyositis
  • Acute viral infections, DNA viral infections (59, 48, 43, 53, 40, 13)
  • Malignant neoplasms (cancers)(40)
  • Alcoholic hepatitis/alcoholic liver disease (32, 48, 40,10,13, 49, 43, 53)
  • Primary sclerosing cholangitis (48, 53)
  • Hepatitis (54)
  • "Sticky" blood (in Africans) (38, 34, 40)
  • Antibodies with a high affinity for polystyrene (used in the test kits)(62, 40, 3)
  • Blood transfusions, multiple blood transfusions (63, 36,13, 49, 43, 41)
  • Multiple myeloma (10, 43, 53)
  • HLA antibodies (to Class I and II leukocyte antigens)(7, 46, 63, 48, 10, 13, 49, 43, 53)
  • Anti-smooth muscle antibody (48)
  • Anti-parietal cell antibody (48)
  • Anti-hepatitis A IgM (antibody)(48)
  • Anti-Hbc IgM (48)
  • Administration of human immunoglobulin preparations pooled before 1985 (10)
  • Haemophilia (10, 49)
  • Haematologic malignant disorders/lymphoma (43, 53, 9, 48, 13)
  • Primary biliary cirrhosis (43, 53, 13, 48)
  • Stevens-Johnson syndrome9, (48, 13)
  • Q-fever with associated hepatitis (61)
  • Heat-treated specimens (51, 57, 24, 49, 48)
  • Lipemic serum (blood with high levels of fat or lipids)(49)
  • Haemolyzed serum (blood where haemoglobin is separated from the red cells)(49)
  • Hyperbilirubinemia (10, 13)
  • Globulins produced during polyclonal gammopathies (which are seen in AIDS risk groups)(10, 13, 48)
  • Healthy individuals as a result of poorly-understood cross-reactions (10)
  • Normal human ribonucleoproteins (48,13)
  • Other retroviruses (8, 55, 14, 48, 13)
  • Anti-mitochondrial antibodies (48, 13)
  • Anti-nuclear antibodies (48, 13, 53)
  • Anti-microsomal antibodies (34)
  • T-cell leukocyte antigen antibodies (48, 13)
  • Proteins on the filter paper (13)
  • Epstein-Barr virus (37)
  • Visceral leishmaniasis (45)
  • Receptive anal sex (39, 64)


1. Agbalika F, Ferchal F, Garnier J-P, et al. 1992. False-positive antigens related to emergence of a 25-30 kD protein detected in organ recipients. AIDS. 6:959-962.

2. Andrade V, Avelleira JC, Marques A, et al. 1991. Leprosy as a cause of false-positive results in serological assays for the detection of antibodies to HIV-1. Intl. J. Leprosy. 59:125.

3. Arnold NL, Slade RA, Jones MM, et al. 1994. Donor follow up of influenza vaccine-related multiple viral enzyme immunoassay reactivity. Vox Sanguinis. 67:191.

4. Ascher D, Roberts C. 1993. Determination of the etiology of seroreversals in HIV testing by antibody fingerprinting. AIDS. 6:241.

5. Barbacid M, Bolgnesi D, Aaronson S. 1980. Humans have antibodies capable of recognizing oncoviral glycoproteins: Demonstration that these antibodies are formed in response to cellular modification of glycoproteins rather than as consequence of exposure to virus. Proc. Natl. Acad. Sci. 77:1617-1621.

6. Biggar R, Melbye M, Sarin P, et al. 1985. ELISA HTLV retrovirus antibody reactivity associated with malaria and immune complexes in healthy Africans. Lancet. ii:520-543.

7. Blanton M, Balakrishnan K, Dumaswala U, et al. 1987. HLA antibodies in blood donors with reactive screening tests for antibody to the immunodeficiency virus. Transfusion. 27(1):118.

8. Blomberg J, Vincic E, Jonsson C, et al. 1990. Identification of regions of HIV-1 p24 reactive with sera which give "indeterminate" results in electrophoretic immunoblots with the help of long synthetic peptides. AIDS Res. Hum. Retro. 6:1363.

9. Burkhardt U, Mertens T, Eggers H. 1987. Comparison of two commercially available anti-HIV ELISA's: Abbott HTLV-III ELA and DuPont HTLV-III ELISA. J. Med. Vir. 23:217.

10. Bylund D, Ziegner U, Hooper D. 1992 Review of testing for human immunodeficiency virus. Clin. Lab. Med. 12:305-333.

11. Challakere K, Rapaport M. 1993. False-positive human immunodeficiency virus type 1 ELISA results in low-risk subjects. West. J. Med. 159(2):214-215.

12. Charmot G, Simon F. 1990. HIV infection and malaria. Revue du practicien. 40:2141.

13. Cordes R, Ryan M. 1995. Pitfalls in HIV testing. Postgraduate Medicine. 98:177.

14. Dock N, Lamberson H, O'Brien T, et al. 1988. Evaluation of atypical human immunodeficiency virus immunoblot reactivity in blood donors. Transfusion. 28:142.

15. Esteva M, Blasini A, Ogly D, et al. 1992. False positive results for antibody to HIV in two men with systemic lupus erythematosus. Ann. Rheum. Dis. 51:1071-1073.

16. Fassbinder W, Kuhni P, Neumayer H. et al. 1986. Prevalence of antibodies against LAV/HTLV-III [HIV] in patients with terminal renal insufficiency treated with hemodialysis and following renal transplantation. Deutsche Medizinische Wochenschrift. 111:1087.

17. Fleming D, Cochi S, Steece R. et al. 1987. Acquired immunodeficiency syndrome in low-incidence areas. JAMA. 258(6):785.

18. Gill MJ, Rachlis A, Anand C. 1991. Five cases of erroneously diagnosed HIV infection. Can. Med. Asso. J. 145(12):1593.

19. Healey D, Bolton W. 1993. Apparent HIV-1 glycoprotein reactivity on Western blot in uninfected blood donors. AIDS. 7:655-658.

20. Hisa J. 1993. False-positive ELISA for human immunodeficiency virus after influenza vaccination. JID. 167:989.

21. Isaacman S. 1989. Positive HIV antibody test results after treatment with hepatitis B immune globulin. JAMA. 262:209.

22. Jackson G, Rubenis M, Knigge M, et al. 1988. Passive immunoneutralisation of human immunodeficiency virus in patients with advanced AIDS. Lancet, Sept. 17:647.

23. Jindal R, Solomon M, Burrows L. 1993. False positive tests for HIV in a woman with lupus and renal failure. NEJM. 328:1281-1282.

24. Jungkind D, DiRenzo S, Young S. 1986. Effect of using heat-inactivated serum with the Abbott human T-cell lymphotropic virus type III [HIV] antibody test. J. Clin. Micro. 23:381.

25. Kashala O, Marlink R, Ilunga M. et al. 1994. Infection with human immunodeficiency virus type 1 (HIV-1) and human T-cell lymphotropic viruses among leprosy patients and contacts: correlation between HIV-1 cross-reactivity and antibodies to lipoarabionomanna. J. Infect. Dis. 169:296-304.

26. Lai-Goldman M, McBride J, Howanitz P, et al. 1987. Presence of HTLV-III [HIV] antibodies in immune serum globulin preparations. Am. J. Clin. Path. 87:635.

27. Langedijk J, Vos W, Doornum G, et al. 1992. Identification of cross-reactive epitopes recognized by HIV-1 false-positive sera. AIDS. 6:1547-1548.

28. Lee D, Eby W, Molinaro G. 1992. HIV false positivity after hepatitis B vaccination. Lancet. 339:1060.

29. Leo-Amador G, Ramirez-Rodriguez J, Galvan-Villegas F, et al. 1990. Antibodies against human immunodeficiency virus in generalized lupus erythematosus. Salud Publica de Mexico. 32:15.

30. Mackenzie W, Davis J, Peterson D. et al. 1992. Multiple false-positive serologic tests for HIV, HTLV-1 and hepatitis C following influenza vaccination, 1991. JAMA. 268:1015-1017.

31. Mathe G. 1992. Is the AIDS virus responsible for the disease? Biomed & Pharmacother. 46:1-2.

32. Mendenhall C, Roselle G, Grossman C, et al. 1986. False-positive tests for HTLV-III [HIV] antibodies in alcoholic patients with hepatitis. NEJM. 314:921.

33. Moore J, Cone E, Alexander S. 1986. HTLV-III [HIV] seropositivity in 1971-1972 parenteral drug abusers - a case of false-positives or evidence of viral exposure? NEJM. 314:1387-1388.

34. Mortimer P, Mortimer J, Parry J. 1985. Which anti-HTLV-III/LAV [HIV] assays for screening and comfirmatory testing? Lancet. Oct. 19, p873.

35. Neale T, Dagger J, Fong R, et al. 1985. False-positive anti-HTLV-III [HIV] serology. New Zealand Med. J. October 23.

36. Ng V. 1991. Serological diagnosis with recombinant peptides/proteins. Clin. Chem. 37:1667-1668.

37. Ozanne G, Fauvel M. 1988. Perfomance and reliability of five commercial enzyme-linked immunosorbent assay kits in screening for anti-human immunodeficiency virus antibody in high-risk subjects. J. Clin. Micro. 26:1496.

38. Papadopulos-Eleopulos E. 1988. Reappraisal of AIDS - Is the oxidation induced by the risk factors the primary cause? Med. Hypo. 25:151.

39. Papadopulos-Eleopulos E, Turner V, and Papadimitriou J. 1993. Is a positive Western blot proof of HIV infection? Bio/Technology. June 11:696-707.

40. Pearlman ES, Ballas SK. 1994. False-positive human immunodeficiency virus screening test related to rabies vaccination. Arch. Pathol. Lab. Med. 118-805.

41. Peternan T, Lang G, Mikos N, et al. Hemodialysis/renal failure. 1986. JAMA. 255:2324.

42. Piszkewicz D. 1987. HTLV-III [HIV] antibodies after immune globulin. JAMA. 257:316.

43. Profitt MR, Yen-Lieberman B. 1993. Laboratory diagnosis of human immunodeficiency virus infection. Inf. Dis. Clin. North Am. 7:203.

44. Ranki A, Kurki P, Reipponen S, et al. 1992. Antibodies to retroviral proteins in autoimmune connective tissue disease. Arthritis and Rheumatism. 35:1483.

45. Ribeiro T, Brites C, Moreira E, et al. 1993. Serologic validation of HIV infection in a tropical area. JAIDS. 6:319.

46. Sayers M, Beatty P, Hansen J. 1986. HLA antibodies as a cause of false-positive reactions in screening enzyme immunoassays for antibodies to human T-lymphotropic virus type III [HIV]. Transfusion. 26(1):114.

47. Sayre KR, Dodd RY, Tegtmeier G, et al. 1996. False-positive human immunodeficiency virus type 1 Western blot tests in non-infected blood donors. Transfusion. 36:45.

48. Schleupner CJ. Detection of HIV-1 infection. In: (Mandell GI, Douglas RG, Bennett JE, eds.) Principles and Practice of Infectious Diseases, 3rd ed. New York: Churchill Livingstone, 1990:1092.

49. Schochetman G, George J. 1992. Serologic tests for the detection of human immunodeficiency virus infection. In AIDS Testing Methodology and Management Issues, Springer-Verlag, New York.

50. Simonsen L, Buffington J, Shapiro C, et al. 1995. Multiple false reactions in viral antibody screening assays after influenza vaccination. Am. J. Epidem. 141-1089.

51. Smith D, Dewhurst S, Shepherd S, et al. 1987. False-positive enzyme-linked immunosorbent assay reactions for antibody to human immunodeficiency virus in a population of midwestern patients with congenital bleeding disorders. Transfusion. 127:112.

52. Snyder H, Fleissner E. 1980. Specificity of human antibodies to oncovirus glycoproteins; Recognition of antigen by natural antibodies directed against carbohydrate structures. Proc. Natl. Acad. Sci. 77:1622-1626.

53. Steckelberg JM, Cockerill F. 1988. Serologic testing for human immunodeficiency virus antibodies. Mayo Clin. Proc. 63:373.

54. Sungar C, Akpolat T, Ozkuyumcu C, et al. Alpha interferon therapy in hemodialysis patients. Nephron. 67:251.

55. Tribe D, Reed D, Lindell P, et al. 1988. Antibodies reactive with human immunodeficiency virus gag-coated antigens (gag reactive only) are a major cause of enzyme-linked immunosorbent assay reactivity in a bood donor population. J. Clin. Micro. April:641.

56. Ujhelyi E, Fust G, Illei G, et al. 1989. Different types of false positive anti-HIV reactions in patients on hemodialysis. Immun. Let. 22:35-40.

57. Van Beers D, Duys M, Maes M, et al. Heat inactivation of serum may interfere with tests for antibodies to LAV/HTLV-III [HIV]. J. Vir. Meth. 12:329.

58. Voevodin A. 1992. HIV screening in Russia. Lancet. 339:1548.

59. Weber B, Moshtaghi-Borojeni M, Brunner M, et al. 1995. Evaluation of the reliability of six current anti-HIV-1/HIV-2 enzyme immunoassays. J. Vir. Meth. 55:97.

60. Wood C, Williams A, McNamara J, et al. 1986. Antibody against the human immunodeficiency virus in commercial intravenous gammaglobulin preparations. Ann. Int. Med. 105:536.

61. Yale S, Degroen P, Tooson J, et al. 1994. Unusual aspects of acute Q fever-associated hepatitis. Mayo Clin. Proc. 69:769.

62. Yoshida T, Matsui T, Kobayashi M, et al. 1987. Evaluation of passive particle agglutination test for antibody to human immunodeficiency virus. J. Clin. Micro. Aug:1433.

63. Yu S, Fong C, Landry M, et al. 1989. A false positive HIV antibody reaction due to transfusion-induced HLA-DR4 sensitization. NEJM.320:1495.

64. National Institue of Justice, AIDS Bulletin. Oct. 1988.