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Day 4 is better than day 3 for a single patch test reading.

Readings were performed on day (D) 2, D3 and D4 after application of patch tests in 88 patients. 90 patch test reactions in 49 patients were interpreted as allergic and of past or present relevance. A single D2 reading detected 58 of the allergic reactions with 32 false-negatives and 23 false-positives. A single D3 reading detected 77 allergic reactions, with 13 false-negatives and 17 false-positives. A single D4 reading detected 85 allergic reactions, with 5 false-negatives and 9 false-positives. Therefore, if only a single reading is feasible, it is better performed on D4 than on D3.

Allergens↗

A study of co-positivity of three multipuncture techniques with intradermal PPD tuberculin.

One hundred thirty-five patients with a reaction of 10 mm or more of induration to 5 TU of intradermal PPD were simultaneously tested with 3 multipuncture techniques to determine the copositivity of the multiple puncture methods with the Mantoux method. Of the 3 multipuncture preparations used (Tine Test, Aplitest, and Mono-Vacc), the Mono-Vacc had 2 (1.5 per cent) false-negative result after 48 hours with only one remaining negative result after 72 hours; whereas the Tine-Test had 21 (15.5 per cent), and the Aplitest had 13 (9.6 per cent) false-negative reactions after 48 hours. Vesicular eruptions were not a commonly noted reaction to the multipuncture preparations. Of the 3 multipuncture preparations commercially available, Mono-Vacc is the preferred reagent because of its significantly lower incidence of false-negative reactions.

Adolescent↗

The sensitivity and specificity of the immediate-spin crossmatch.

The immediate-spin (IS) crossmatch is used to detect ABO incompatibility between donor red cells (RBCs) and the serum of the intended recipient. However, this test may be positive in the absence of ABO incompatibility (false positive) or it may be negative when ABO incompatibility exists (false negative). During a 25-month study, the rates of both false-positive and false-negative IS crossmatch results were evaluated, and the sensitivity and specificity of the IS crossmatch were determined. During the study period, 53,656 IS crossmatches were performed for patients without significant RBC antibodies. Fifty-five patients had positive IS crossmatches, and no false-negative reactions were found. In tests of 55 patients with positive IS crossmatches, 77 false-positive and 5 true-positive reactions were noted. The causes of the false-positive reactions were rouleaux (36 patients), cold-reactive antibodies (8 patients), a combination of rouleaux and cold-reactive antibodies (2 patients), fibrin clot (1 patient), and undetermined (3 patients). The sensitivity and specificity of the IS crossmatch were 100 and 99.86 percent, respectively. Laboratory personnel should be aware that the IS crossmatch may have false-positive or false-negative results, and they should develop written protocols to distinguish quickly between true-positive and false-positive reactions.

ABO Blood-Group System↗

Development of a novel internal positive control for Taqman based assays.

Development of rapid amplification assays for the detection and identification of biological threat agents has become a focus of diagnostic efforts in recent years. The use of real-time PCR assays as diagnostic tools depends upon two critical processes. First, nucleic acid purification must provide template that is both amplifiable and free of PCR inhibitors. Second, the assays themselves must be sensitive and specific for their nucleic acid targets. A differentiation must be made between results achieved due to the lack of target nucleic acid (true negatives) and those produced due to the inability to amplify target DNA (false negatives) so confidence in negative reactions is possible. False negatives can occur when inhibitors are present in the sample being tested, especially if clinical samples such as blood are analyzed. To address the problem of detecting inhibition in purified nucleic acids, an exogenous internal positive control (IPC) based on Taqman chemistry was developed. A previously optimized assay was cloned and the primer and probe sites were mutated to produce novel sequences with no known homology to published sequence data. The IPC was sensitive to a variety of inhibitors, including hemoglobin, heparin, EDTA, humic acids, and fulvic acid. It was also equally sensitive to inhibition when labeled with either 6FAM or ROX dyes. In addition, the IPC was successfully multiplexed with agent specific assays without any loss in their sensitivity. The designed IPC assay has proven to be an effective tool for monitoring inhibitors of PCR and builds confidence in negative results obtained with agent specific assays.

False Negative Reactions↗

Tuberculin skin testing: present status.

The incidence of tuberculosis is on the rise, with a reported 18% increase between 1985 and 1992. There is every reason to think that this trend will continue in the 1990s. Expectedly, there is a renewed interest in early detection and prevention of this potentially life threatening disease. Tuberculin skin testing remains a valuable diagnostic tool for the detection, control, and elimination of tuberculosis. It involves the intracutaneous injection of culture extracts of tubercle bacilli as a classic example of delayed hypersensitivity reaction. Despite proper technique and interpretation of the test, its usefulness is limited by cross-reaction caused by other mycobacterial antigens. False-negative reaction is a common confounding problem. It is therefore important to interpret the test based on the overall clinical context of the individual being tested. This article aims to review the basic immunology, proper technique, interpretation, and indication for the test.

Aged↗

[Automatic reading of ABO and Rh groups on microplates using FMC medium and an IBG Systems reader].

We present in this paper our experience in the routine use of an automatic reader for microtiter plates (IBG Systems). A total of 2044 samples from blood donors have been tested for ABO (haematic and seric) and Rh (D antigen) blood group typing. The red blood cell samples have been tested against monoclonal anti-A, anti-B, anti-AB and anti-Rh 1 (D) sera (using two different anti-D reagents). As a negative control, 3% albumin solution was employed. In order to determine the seric groups, the plasma samples were tested against A1, A2, B and 0 cells. In all cases the red cells were suspended in Ficoll 400R-Methyl cellulose (FMC), and 0.01 Bromelin was added to red blood cell samples to be typed. The obstacles in the automatic readings were mainly solved by visual reading of the microplates. In the 2,044 samples analysed with the automatic reader, 19 discrepancies (0.94%) were found in ABO typing. In all cases the error was in the seric blood group. Three false positive reactions were found with 0 red cells. Two false negative with B red cells, interpreted by the automatic reader as doubtful results. Three positive readings were detected, in one case the positivity was due to alloantibodies (C + D specificities), the other two cases were autoagglutinations. False negative reactions were found, in 11 cases, 8 of them due to lipaemia and the remaining three were haemolysed plasma samples. It should be stressed that out of 19 cases of discrepancy, only 5 (0.24%) were due to the automatic reader while the remaining were due, to sample troubles.(ABSTRACT TRUNCATED AT 250 WORDS)

ABO Blood-Group System↗

Contact sensitivity to nickel and other metals in jewelry reactors.

BACKGROUND: Many patients who give a history of a dermatitis reaction to jewelry or metal contact with skin are negative to metals on standard patch testing. Some may be showing false-negative reactions. OBJECTIVE: Our purpose was to determine whether patients with a history of jewelry reactions but whose standard patch tests were negative have a false-negative reaction or are allergic to metals other than nickel, cobalt, or chromium. METHODS: Four hundred forty-nine patients were studied who gave a history of reacting to jewelry or metal. Of these, 210 were tested to the metals in the European standard series (ie, nickel, cobalt, and chromate), and 239 were tested to the metals in the standard series and to an extended metal series of palladium, gold, platinum, a second nickel salt, and a nickel/cobalt mixture. These were compared with 752 patients who did not give a history of jewelry or metal reactions, of which 50, besides the standard series, were also treated with the additional metal series. RESULTS: A higher proportion of jewelry-reactive patients tested with the extended series reacted to nickel (and to other metals) than those who were tested only with the European standard series: (61% vs 38%; P <.0001 ). The use of the extended series showed that palladium allergy was common, present in 34% of nickel-allergic patients, but it always occurred with nickel sensitivity. Gold allergy coexisted with nickel sensitivity in 10% of cases. Testing simultaneously with separate patches containing 5% nickel sulfate and 5% nickel chloride showed a concordance of 71% in identified nickel-sensitive patients. Nickel sulfate was more likely than nickel chloride to detect nickel sensitivity. The use of a combined preparation of 2.5% nickel sulfate and 0.5% cobalt chloride in petrolatum revealed only 3 jewelry-reactive patients who were negative to other metals. There was a slightly higher proportion of atopic patients in the patch test-negative jewelry reactors group than in the positive group; however, the difference was not significant and it was not sufficient to account for the negative findings. CONCLUSION: Some jewelry reactors who had negative patch tests are likely to be subclinically allergic to nickel. We suggest that the higher number of antigens, or perhaps the larger nickel load, in the extended metal series, resulted in a larger proportion of patients reacting. To better demonstrate nickel allergy in jewelry reactors, patients should be patch tested to a metal series that contains palladium and gold salts and perhaps a second nickel patch because these may reveal the presence of nickel sensitivity when standard patch tests would otherwise have been negative.

Chromium↗

Assessment of an enzyme-linked immunosorbent assay using a Taenia hydatigena fraction antigen in the diagnosis of cysticercosis in cattle.

Enzyme linked immunosorbent assay (ELISA) using a fraction of larval Taenia hydatigena cyst fluid antigen was carried out on 469 bovine sera collected at slaughter from feedlot cattle for the presence of anticysticercosis antibodies. Cysticerci, in low numbers, were found in the heart, tongue and/or masseter muscles of 84 of the 469 cattle at postmortem inspection. Only nine sera gave positive ELISA reactions and in only one of these nine animals were cysticerci found. Within the limitations of this study, the high rate of false negative and false positive reactions suggests that the ELISA with the antigen used is not a satisfactory procedure to diagnose cysticercosis in cattle, at least in animals with light infections.

Animals↗

[False reactions in hydatic serology in order to improve its value (author's transl)].

We have studied false reactions in hydatic serology in order to improve its value. We have found false positive reactions in two cases of tumor (hepatic sclerocholangiomatosis and pleural mesothelioma) and among children showing cor's milk protein intolerance when serum is tested with an antigen extracted from ox's lung. As for false negative reactions, they are scarcely due to circulating antigens. We have found, for 34 falsely negative serums, IgG, IgM and above all IgE levels lower than the levels of patients showing a positive serology.

Adolescent↗

Evaluation of four qualitative methods for detection of beta-lactamase production in Staphylococcus and Micrococcus species.

Four qualitative methods for the detection of beta-lactamase production in Staphylococcus and Micrococcus species were evaluated and compared with a quantitative macroiodometric reference method. The disc diffusion test with penicillin G and the cloverleaf method could not separate beta-lactamase-positive from beta-lactamase-negative strains. Two applications of the chromogenic cephalosporin test, using uninduced strains and strains grown on blood agar plates, gave a large number of false negative and false positive results. False negative reactions were most common among uninduced strains, while the false positive reactions were most often recorded for Staphylococcus saprophyticus. A high degree of efficiency was recorded for the nitrocefin spot test, using induced strains grown on antibiotic susceptibility agar, and for the starch-iodine plate method. The starch-iodine plate with methicillin as inducer gave the most reliable results.

Bacteriological Techniques↗

Comparison of three commercial cryptococcal latex kits for detection of cryptococcal antigen.

Three commercial latex kits, IBL, MYCO-Immune, and IMMY, for the detection of cryptococcal antigen were compared in regard to sensitivity, specificity, and height of antigen titers. A total of 218 cerebrospinal fluid and 79 serum specimens from 239 patients were included. Twenty-two patients had culture-proven disseminated cryptococcosis. Both the IBL and MYCO-Immune kits had sensitivities of 100%, and the IMMY kit had sensitivities of 82.6 and 45.4% in CSF and serum specimens, respectively. There was one false-positive reaction in serum with the MYCO-Immune kit and one false-negative reaction on screen only with all three kits. Rheumatoid factor-containing sera were used to check the agglutination titers between matching anti-cryptococcal globulin reagents and normal globulin reagents. The finding that agglutination titer with anti-cryptococcal globulin reagents was fourfold higher than with normal globulin reagents in the MYCO-Immune kit is considered to be a cause for a false-positive reaction in serum.

Antigens, Bacterial↗

Red cell phenotyping using hexadimethrine bromide (Polybrene) in a microplate system.

To provide rapidly phenotyped units of blood, we adapted the hexadimethrine bromide (Polybrene) technique to microplate technology. Pilot samples from 282 donor units were phenotyped for antigens in the Rh, Kidd, Kell, Duffy, and Ss systems with a standard tube-testing method and a Polybrene-microplate (P-MP) technique. Diluted antisera and a 1 percent red cell suspension were used to give P-MP reactions that were accurate and easy to interpret. One microplate, containing 96 tests, was prepared and read within 15 minutes for P-MP tests yielding direct agglutination (Rh, Jkb, Fya, Fyb), or within 19 minutes for P-MP tests requiring an antiglobulin phase (Jka, K, S, s). No false-positive results were found. No false-negative reactions were found in typing for Rh, K, S, s, or Jka antigens. Phenotyping for Jkb and Duffy antigens gave a false-negative rate of less than 0.015. Considerable savings in reagents were obtained through microplate miniaturization and through the enhancement of apparent antibody avidity in the Polybrene/low ionic medium which permitted dilution of reagent antisera. The P-MP technique affords a rapid, accurate, simple, and inexpensive means of phenotyping large numbers of donor units.

Blood Group Antigens↗

Methyldibromoglutaronitrile is an important contact allergen in The Netherlands.

From 15 May to 15 December 1994, 2943 patients suspected of having contact dermatitis (1955 women, 988 men) were patch tested with methyldibromoglutaronitrile 0.3%, 0.1% and 0.05% pet. 119 patients (4.0%; women 4.1%, men 3.8%) proved to be allergic. 71% of the reactions were considered to be relevant. In 2/3 of the patients, causative products were cosmetics, in 1/3 moistened toilet tissues. Testing with methyldibromoglutaronitrile at lower concentrations (0.05% and 0.1%) and with commercial allergens (Euxyl K 400 and methyldibromoglutaronitrile, both containing methyldibromoglutaronitrile 0.1%), resulted in a number of false-negative reactions. All preservatives in the European standard series had lower scores than the 4% positive reactions to methyldibromoglutaronitrile (formaldehyde 2.0%, MCI/MI (Kathon CG) 3.2%, parabens 1.0%, quaternium-15 1.3%). It is concluded that methyldibromoglutaronitrile (present in the commercial preservative Euxyl K 400) is an important contact allergen in the Netherlands in cosmetics and moistened toilet tissues. It should be added to cosmetics series and to proctological series. The optimal test concentration is unknow, but may be 0.3% pet. The concentration of 0.1% methyldibromoglutaronitrile in the currently available commercial allergens appears to be too low, resulting in a number of false-negative reactions.

Adolescent↗

Cutaneous adverse reactions to clindamycin: results of skin tests and oral exposure.

BACKGROUND: Clindamycin is an antibiotic used in anaerobic and severe complicated infections. It is often selected for patients with a history of allergy to other antibiotics. OBJECTIVES: To study the occurrence of clindamycin hypersensitivity and to determine whether skin tests are useful in cases of suspected clindamycin allergy. METHODS: Six patients with an exanthematous rash and a history strongly suggestive of clindamycin hypersensitivity were studied with skin tests and oral exposure. Cases of suspected adverse drug reactions to clindamycin reported to the National Register of Adverse Effects of Drugs (NRAED) in Finland during 1973-2000 were analysed. RESULTS: In the skin tests true-positive patch test reactions were seen in four of six patients, while 22 healthy control patients were negative. One false-positive and one false-negative patch test reaction were seen. During 1973-2000, 29 suspected cases of skin and/or mucosal membranes affected by clindamycin were reported to the NRAED. CONCLUSIONS: Clindamycin hypersensitivity is not common. Delayed-type allergic reactions occur and patch tests are useful in those cases. Oral exposure is the method of choice if possible, as false-negative and false-positive reactions may occur.

Administration, Oral↗

False-negative syphilis screening due to change in temperature.

BACKGROUND AND OBJECTIVES: Inaccurate test results for syphilis may cause an individual to experience serious effects. GOAL OF THE STUDY: Investigate potential sources of error and test limitations causing false-negative reactions. STUDY DESIGN: In 5 months, two laboratories screened 2,232 patients for syphilis by the Rapid Plasma Reagin (RPR) test. RESULTS: The hospital laboratory reported 5.3% (64/1,210) of patients' test as reactive on initial screening, and the research laboratory found 6.4% (78/1,210) reactive. Fourteen reactive patients were incorrectly reported negative by the hospital laboratory, as confirmed by both laboratories. A refrigerated centrifuge in the hospital laboratory possibly caused sera to be cooled before testing, producing false-negative results. When its temperature was adjusted from 4 degrees C to 27 degrees C, an additional 1,022 samples tested were consistent between the two laboratories. CONCLUSION: Cold temperature produces false-negative reactions for syphilis screenings in patients' samples with titers < 1:4 dilution. Patients' samples with titers > or = 1:16 dilution were not affected. According to this study, incorrect temperatures for test sera can alter testing outcomes. Therefore, test manufacturer's directions must be strictly followed.

Bias↗

Comparative evaluation of identification systems for testing methicillin-resistant strains of Staphylococcus aureus.

Several commercial systems are available to distinguish between Staphylococcus aureus and the coagulase-negative species of the Micrococcaceae family. Four latex agglutination systems (Accu-Staph, SeroSTAT, Staphaurex, and Staphylatex) and two hemagglutination systems (Hemastaph and Staphyloslide) were compared for their performance in the rapid identification of 232 isolates of staphylococci, including 114 of methicillin-resistant S. aureus. Accu-Staph, Staphaurex, and Staphyloslide correctly identified 100% of the methicillin-resistant S. aureus isolates; Hemastaph and Staphylatex, 99.1%; and SeroSTAT, 94.7%. Most reactions were easy to interpret, although 15% of the SeroSTAT reactions were weak. Autoagglutination occurred only with isolates of coagulase-negative staphylococci. False-positive reactions were rare and occurred only with systems which did not detect autoagglutination. Five of these six systems appear to be adequate for the rapid identification of S. aureus, including methicillin-resistant isolates.

Coagulase↗

Detection of antibody in Aleutian disease of mink: comparison of enzyme-linked immunosorbent assay and counterimmunoelectrophoresis.

Experiments were undertaken to investigate the potential of the enzyme-linked immunosorbent assay (ELISA) as a screening test for the diagnosis of the 2 known naturally occurring forms of Aleutian disease of mink. Anti-Aleutian disease virus (ADV) antibody activity was not detectable in the sera of mink with nonprogressive Aleutian disease despite the demonstration of antibody by counterimmunoelectrophoresis (CIEP) in the same sera. Anti-ADV antibody was detectable in 93% of sera from mink at various stages of experimentally induced progressive Aleutian disease. False-negative reactions occurred in sera which demonstrated high anti-ADV antibody titers by CIEP. As a consequence of the high prevalence of false-negative reactions, the ELISA was not considered to be an effective screening test. However, using CIEP as an indicator of ADV infection, the ELISA may be useful in differentiating mink with nonprogressive Aleutian disease from mink with progressive Aleutian disease.

Aleutian Mink Disease↗

Urine protein determination in dogs and cats: comparison of dipstick and sulfasalicylic acid procedures.

Protein levels in urine specimens from 91 dogs and 65 cats were evaluated by sulfasalicylic acid precipitation (SSA) and dipstick methods. The dipstick frequently yielded reactions for protein that were greater than the level of protein indicated by SSA (i.e., false positive reactions), although no false negative reactions for protein were noted. All urine specimens with protein levels equal to or greater than 100 mg/dl by SSA had dipstick results of 3 +. Results of this study suggest that dipstick analysis for urine protein is an adequate screening procedure for the selection of urines for quantitative analysis of protein and creatinine to assess proteinuria.

Journal Article↗