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To answer the question of whether proximity of Multi-Test skin test sites causes false positive reactions, 50 subjects with prior 4+ skin test responses were retested with antigens and ten additional subjects were tested with histamine (1.8 mg/mL). No positive readings occurred at any of the replicate saline control sites adjacent to 3+ or 4+ allergen sites or to 3+ histamine reactions.
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Acute and chronic biological false positive reactions to serological tests for syphilis are discussed. Three cases are presented, one showing an acute and one a chronic reaction, and one in which it was not possible to determine if the reaction was acute or chronic. Once a biological false positive reaction is found, serum from the patient should be tested at intervals for at least six months to determine if the reaction is acute or chronic. If a chronic reaction is found the patient should then be investigated to find the underlying cause. Problem sera from 753 patients are reviewed. Biological false positive reactions were shown by 47 sera and 257 sera gave results indicating they came from patients infected with syphilis. The reactive sera were divided into four groups: sera from blood donors, antenatal patients, cases of recent vaccination, and a miscellaneous group. It was found that in the first three groups the number of sera from patients showing biological false positive reactions was equalled or exceeded by the number showing syphilitic reactions. Biological false positive reactions in sera from cases of lepromatous leprosy and in sera from the inhabitants of the Okapa region of New Guinea are also discussed. It is concluded that sera which give reactive results to reagin detection tests for syphilis and come from patients known to have conditions which cause acute or chronic biological false positive reactions should never be dismissed as such. Experience has shown that a number of these sera will give reactive Treponema pallidum immobilization test results, that is, they come from patients who have or have had syphilis.
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The frequency of biologically false positive reactions to serological tests for syphilis depends on a number of factors, including the individual immunological response, the number and type of serological tests performed, and the stage of the disease producing such reactions; the relative importance of such factors is discussed. The author also considers in detail the diseases or conditions giving rise to acute or chronic biologically false positive reactions.A variety of verification tests exists for differentiating the true syphilitic reaction from the biologically false positive reaction, but none is so accurate as the Treponema pallidum immobilization and immune adherence tests, which the author considers should be used when others have proved inconclusive. In the final section of his paper, he indicates the steps to be followed in attempting to distinguish between latent syphilis and biologically false positive reactions in persons with positive serological tests but no anamnestic or clinical evidence of syphilis.
There are limited data about false-positive reactions against hepatitis C virus (HCV) in syphilitic patients and false-positive reactions against syphilis in the patients with HCV infection. The aim of this study was to demonstrate the false-positivity of syphilis in patients with HCV infection, the false-positivity of anti-HCV in patients with syphilis and the validity of the serological tests in such patients. Fifty patients with positive anti-HCV, 21 patients with positive VDRL and 50 healthy subjects were studied. Syphilis serology was determined by the Venereal Disease Research Laboratory (VDRL) test and microhemagglutination for T. pallidum (MHA-TP) test. Hepatitis C serology was determined by a second generation ELISA (Ortho Diagnostics) test for HCV antibody, and anti-HCV positive patients were tested for HCV RNA by polymerase chain reaction (PCR). All assays were performed on all subjects. Not only the false-positive VDRL reaction in the patients with HCV infection but also false-positive anti-HCV tests in syphilitic patients have been observed. Four patients with syphilis had positive anti-HCV and negative HCV-RNA, whereas 10% (5 of 50) of patients with hepatitis C infection had positive VDRL and these patients were negative for MHA-TP test. The rates of false-positivity of VDRL and anti-HCV were higher than within the control group (p < 0.05). According to these data, positive anti-HCV in syphilitic patients and positive VDRL in chronic hepatitis C may be false-positive results with regard to the reaginic tests. Therefore, therapeutic measures should not be initiated without confirmation with a treponemal test or PCR. VDRL and HCV-ELISA tests may be interacted with IgM or IgG antibodies. This relationship should be investigated in further studies.
False-positive reactions were observed in commercial test kits designed to detect FeLV infections in cats. The false-positive reactions were associated with heterophilic antibody in cat serum directed primarily against mouse immunoglobulins. Of 2,830 sera tested, 579 were considered FeLV positive or equivocal via ELISA; of these, 10 were found that did not have FeLV antigen but did have anti-mouse antibody. With 95% confidence, 0.14% to 0.57% of cats would have such false-positive reactions in ELISA, because one in 283 (0.35%) cats had anti-mouse antibodies.
Problem sera from 375 blood donors were investigated for biological false-positive reactions to serological tests for syphilis. Problem sera are those sera in which submitting laboratories have found a reactive result to a screening test for syphilis. On each serum a cardiolipin Wassermann reaction, a Venereal Disease Research Laboratory test, a Reiter protein-complement fixation test, a Treponema pallidum immobilization test, and a fluorescent treponemal antibody absorption test were performed. Of the sera 49.9% were found to be non-reactive in all five tests, 28.8% gave results indicating a diagnosis of syphilis, and 21.3% showed biological false-positive reactions. There were 80 sera from blood donors which gave biological false-positive reactions. A further specimen of serum from 67 of these donors was tested after an interval of a minimum of six months. Sixty-six of the sera showed chronic biological false-positive reactions. Some donors were only found reactive for the first time after they had given very many donations of blood. It is concluded that sudden blood loss, as in blood donation, appears to stimulate the production of excess reagin in certain individuals, causing a chronic biological false positive reaction to serological tests for syphilis. This may only appear after several blood donations have been made.
We investigated false-positive reactions obtained from a drug screening test using a Triage panel. We detected 2 cases giving false-positive reaction for AMP (amphetamine, methamphetamine) during the screening of 187 normal subjects. Subsequent follow up testing by high-performance liquid chromatography (HPLC), showed both to be false-positive reactions. As both cases have a history of ingesting the herbal drug, Ma-huang (Ephedra sinica (Ephedraceae)), containing ephedrine, we examined the relationship between false-positive reactions on Triage and Ma-huang. All urine samples collected from 7 healthy volunteers following administration of Ma-huang indicated AMP positive on Triage. Also a high ratio of AMP positives was observed in the patients who were administered Ma-huang-containing drugs at the hospital. However, none of them were identified as true-positives by HPLC or gas chromatography mass spectrometry (GC/MS) analysis. The extract of Ma-huang contained in herbal drugs, which otherwise contain neither amphetamine nor its derivatives, gives (AMP) positive indications on Triage. We speculate that unidentified components of Ma-huang cause the false-positive reactions. We suggest that follow-up tests by GC/MS or HPLC are needed wherever a positive result is obtained from a screening test by Triage. Furthermore, it will be established to continue collecting information on prescribed and non-prescribed drugs.
AIM: The Captia Syph G enzyme immuno assay (EAI) offers the potential for the rapid automated detection of syphilis antibodies. This study was designed to assess the role of other sexually transmitted diseases (STDs) in producing false positive reactions in the Captia Syph G EIA. The role of rheumatoid factor (RF) as a potential source of false positives was also analysed. METHODS: Patients who attended a genitourinary medicine (GUM) department and gave a false positive reaction with the EIA between 1988 and 1990 were compared with women undergoing antenatal testing and with the control clinic population (EIA negative) over the same time period. The incidence of sexually transmitted disease (STD) in the clinic population and the false positive reactors was measured in relation to gonorrhoea, chlamydia, genital warts, candidiasis, "other conditions not requiring treatment" and "other conditions requiring treatment." Male: female sex ratios were also compared. Ninety two RF positive sera were analysed with the EIA. RESULTS: The rate of false positive reactions did not differ with respect to the diagnosis within the GUM clinic population. The antenatal group of women, however, had a lower incidence of false positive reactions than the GUM clinic group. No RF positive sera were positive on Captia Syph G EIA testing. CONCLUSIONS: There is no cross reaction between Captia Syph G EIA and any specific STD or with RF positive sera. The lower incidence of false positive reactions in antenatal women is unexplained but may be related to physiological changes associated with pregnancy.
Paget cells from cases of mammary and extramammary Paget's disease were examined for carcinoembryonic antigen (CEA) and CEA-related antigens by the immunoperoxidase method. Paget cells showed a conspicuous positive reaction with antiserum to CEA, but were negative when nonspecific cross-reacting-antigen (NCA)-absorbed antiserum to CEA, or a monoclonal antibody to CEA was used as the detecting agents. Paget cells may contain large amounts of NCA antigen or CEA-related substances.
A solid-phase enzyme-linked immunosorbent assay (ELISA), based on the "sandwich" principle with use of microtiter plates, was developed for the detection of hepatitis B surface antigen (HBSAg). Results could be read within one day by the naked eye or by colorimeter. The detection level was less than or equal to 5-10 ng of HBSAg/ml. The sensitivities of ELISA and radioimmunoassay were about the same in dilution series and in a follow-up study of 19 patients with acute hepatitis B infection. In 11 European medical centers where greater than 50,000 samples were tested, ELISA detected significantly more HBSAg-positive samples than a reversed hemagglutinatiom test. No significant difference in sensitivity between ELISA and radioimmunoassay could be demonstrated. On the average, 2.2% of readings were false-positive reactions. Falsely positive samples were identified by a confirmatory test.
A sandwich ELISA for quantification of the endometrial protein PP14 revealed false positive reactions in 81% of male sera (n = 54). The PP14 ELISA was based on two monoclonal antibodies (Mabs) with different epitope specificities--a catcher and a biotinylated indicator. The monoclonal antibodies were purified by protein G affinity chromatography from culture supernatant containing 10% (v/v) fetal calf serum (FCS). Human anti-animal IgG (bovine, mouse, horse, and swine) antibodies and human anti-bovine serum albumin antibodies were measured using an ELISA design, with direct bridging of the solid phase and biotinylated antigens. The false positive reactions were abolished by addition of 1% (v/v) bovine serum to the dilution buffer (DB). Human anti-bovine IgG antibodies (HABIA) were detected in 99 out of 104 sera from blood donors (50 females; 54 males). HABIA levels in male sera (n = 54) were positively correlated to the false positive signals in the PP14 ELISA (r = 0.923; p < 0.0001). Antibodies to IgG from other mammalian species (mouse, horse, and swine) were also detected in the donor sera, but levels and frequencies were lower compared to that of HABIA. Furthermore, HABIA were positively correlated to human anti-bovine serum albumin antibodies in the donor sera (r = 0.639; p < 0.0001; n = 103). HABIA (prevalence 95%) cause false positive reactions due to crossbinding of contaminating bovine IgG and/or crossreaction with mouse IgG in two-site immunoassays. The apparent presence of human anti-mouse IgG antibodies (HAMA), described to create false positive results, may be due to a crossreacting fraction of the polyclonal circulating antibodies against bovine IgG.
False positive reactions to the fluorescence test (FTA-ABS) were found in a group of dermatological patients compared to another group, made up of healthy subjects and blood-donors. Reactivity to FTA-ABS test was particularly evident in patients suffering from Herpes simplex genitalis, allergic diseases and collagenoses. Practical and theoretical aspects of this finding are discussed.
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False-positive reactions in the Venereal Disease Research Laboratory test are well known, whereas a positive fluorescent treponemal antibody absorption assay is rarely thought to be a false positive. The non-recognition of serological false-positive tests for syphilis may have negative prognostic and social implications.
Anti-M2 of anti-mitochondrial antibodies is recognized as the specific autoantibody detected in sera from patients with primary biliary cirrhosis (PBC). The IgG class and IgM class of this antibody can be separately measured using each ELISA. In the present study, false positive reactions were found in some sera from non-PBC patients such as acute hepatitis A, syphilis and rheumatoid arthritis using the IgM anti-M2 ELISA. They showed an increase of polyclonal IgM, and positivity for IgM anti-cardiolipin or rheumatoid factors, respectively. So, we developed a means to prevent these false positive reactions. First, dilutions of test sera at 1:1000-fold were carried out in addition to the original method at 1:100-fold. Secondly, some blocking reagents were added into the buffer system. By serum dilution, non-specific bindings disappeared in most samples other than showing an increase in polyclonal IgM. Moreover, the addition of suitable blocking reagents such as fetal bovine serum (FBS) and skimmed milk into the buffer system could prevent these non-specific bindings. From these findings, the procedure of optical serum dilution and the addition of suitable blocking reagents successfully prevented false positive reactions in this IgM anti-M2 ELISA.
Specificity of the Dot-enzyme-linked immunosorbent assay (Dot-ELISA) for visceral leishmaniasis was significantly improved through the use of enzyme-conjugated antisera specific for IgG heavy chains. Of sera from Kenyans with visceral leishmaniasis, 97% (29/30) were positive using horseradish peroxidase (HRP)-conjugated anti-IgG (heavy and light chain specific) which detected bound IgG and IgM. False positive reactions occurred in 80% of sera from both trypanosomiasis-infected patients (8/10) and apparently healthy Africans (24/30). HRP-conjugated anti-IgG (heavy chain specific), which detected only bound IgG, significantly reduced false positive reactions among trypanosomiasis-infected (2/10, P less than 0.02) and healthy Africans (6/30, P less than 0.001), without reducing test sensitivity in leishmaniasis patients. No false positives occurred when either HRP-conjugated antiserum was used to assay sera from 30 North Americans. Application of enzyme-conjugated antisera specific for IgG improves the serodiagnostic value of the Dot-ELISA for individual patient evaluation and epidemiologic investigations.