Giant tuberculin reaction associated with the homeopathic drug tuberculinum: a case report.

Giant reactions to the tuberculin skin test are extremely rare and have been previously reported almost exclusively in patients with lepromatous leprosy. We herein report a giant tuberculin reaction associated with the homeopathic drug Tuberculinum in a patient with no evidence of active tuberculosis or leprosy.

Creating falseness-How to establish statistical evidence of the untrue.

Null hypothesis significance testing is the typical statistical approach in search of the truthfulness of hypotheses. This method does not formally consider the prior credence in the hypothesis, which affects the chances of reaching correct conclusions. When scientifically implausible or empirically weakly supported hypotheses are tested, there is an increased risk that a positive finding in a test in fact is false positive. This article argues that when scientifically weakly supported hypotheses are tested repeatedly-such as when studying the clinical effects of homeopathy-the accumulation of false positive study findings will risk providing false evidence also in systematic reviews and meta-analyses. False positive findings are detrimental to science and society, as once published, they accumulate persistent untrue evidence, which risks giving rise to nonpurposive research programmes, policy changes, and promotion of ineffective treatments. The problems with false positive findings are discussed, and advice is given on how to minimize the problem. The standard of evidence of a hypothesis should depend not only on the results of statistical analyses but also on its a priori support. Positive findings from studies investigating hypotheses with poor theoretical and empirical foundations should be viewed as tentative until the results are replicated and/or the hypothesis gains more empirical evidence supporting it as likely to be true.

Effects of homeopathic mother tinctures on breath alcohol testing.

In some countries, it is illegal to drive with any detectable amount of alcohol in blood; in others, the legal limit is 0.5 g/L or lower. Recently, some defendants charged with driving under the influence of alcohol and have claimed that positive breath alcohol test results were due to the ingestion of homeopathic mother tinctures. These preparations are obtained by maceration, digestion, infusion, or decoction of herbal material in hydroalcoholic solvent. A series of tests were conducted to evaluate the alcoholic content of three homeopathic mother tinctures and their ability to produce inaccurate breath alcohol results. Nine of 30 subjects gave positive results (0.11-0.82 g/L) when tests were taken within 1 min after drinking mother tincture. All tests taken at least 15 min after the mother tincture consumption and resulted in alcohol-free readings. An observation period of 15-20 min prior to breath alcohol testing eliminates the possibility of false-positive results.

Evaluation of the automated Bact-Alert system for pediatric blood culturing.

The Organon Teknika BacT/Alert (Organon Teknika, Durham, NC), using the Pedi-BacT 20 mL aerobic bottle (BPBCS) was compared to the Wampole Isolator (WI) 1.5 Microbial tube (Wampole Laboratories, Cranbury, NJ), for detection and recovery of pediatric pathogens. The BPBCS continuously monitors culture bottles for changes in CO2 concentrations, while WI cultures are examined twice daily for appearance of colonial growth on agar media. Of 5,175 paired blood cultures, 383 pathogens were recovered from 606 positive cultures. There were 272 pathogens recovered by both systems, 64 from BPBCS only, and 47 from WI only. Overall recovery rates were 88% for BPBCS and 83% for WI. There was no significant difference between the two systems in detection or times to positivity of staphylococci, Enterobacteriaceae, or pseudomonads. Trends toward better recovery of streptococci (20 vs. 10) and fastidious microaerophiles (3 vs. 0) were found with BPBCS, whereas more slowly growing pathogens (Rochalimaea henselae [1], Mycobacterium avium-intracellulare [1]) were recovered by WI only, but because of their lower frequency did not achieve statistical significance. Detection of Haemophilus influenzae (14.9 hours in WI vs. 45.4 hours in BPBCS) was faster with WI. False positive plus contaminant cultures were detected in 5.9% BPBCS versus 1.5% WI. BPBCS offers detection of bacteremia at a rate comparable to WI with advantages of automation.

Sensitivity and specificity of commercial ELISA kits for screening anti-LAV/HTLV III.

Two panels consisting of 236 and 258 anti-LAV/HTLV III positive and 64 and 50 negative sera, respectively (determined in Western blots), were tested with 8 different, commercially available ELISA test kits for detecting LAV/HTLV III antibodies (Abbott, Dupont, Electro-Nucleonics, Litton, Organon, Pasteur, Sorin, Wellcome). Positive sera were selected for levels of antibodies ranging from average to very high, and the negative sera included both negative sera and sera with antibodies to cellular components such as HLA antigens or nuclear membranes. In examination of the 2 panels, the sensitivity of the tests ranged from 97.2 to 100%, and the specificity from 70.0 to 100%. No test was ideal.

Development of enzyme-linked immunosorbent assay for hepatitis B e antigen.

Hepatitis B e antigen (HBeAg) in sera indicates infectivity and when found during pregnancy, indicates a need for vaccination against hepatitis B virus. A sensitive test for HBeAg is needed in all hospitals but this test is expensive. Local development of enzyme-linked immunosorbent assay (ELISA) for HBeAg and its antibody (anti-HBe) was considered necessary and it was successfully conducted. The developed test was compared with ELISA test for HBeAg and anti-HBe manufactured by Organon Teknika (205 routine specimens and 103 sera positive for HBsAg) and Roche Diagnostic (160 routine specimens). The locally made and imported kits showed overall agreement of 97.5 to 98.1 per cent and the locally made test was always slightly more sensitive. The local test was also rapid, reproducible, and specific. The development lead to self reliance on ELISA test for HBeAg and anti-HBe.

Laboratory methods for quantitating HIV RNA.

AIDS: High plasma HIV RNA levels have been shown to be a strong predictor of rapid progression to AIDS after HIV seroconversion. Recently, three quantitation assays that are less expensive, easier to perform, and more reproducible than older methods were introduced: 1) Amplicor HIV Monitor Test (Roche Molecular Systems); 2) Nucleic Acid Sequence-Based Amplification (NASBA) (Organon Technika); and 3) Quantiplex HIV RNA assay, a branched DNA (bDNA) technology (Chiron Corporation). The Amplicor HIV Monitor Test requires 200 uL plasma samples for reverse transcription, polymerase chain reaction (PCR) and nonradioactive detection of amplified DNA. It has a three-log dynamic range with a lower limit of sensitivity of about 500 copies of HIV RNA/mL. The NASBA assay uses nonradioactive detection to measure RNA after nucleic acid isolation and amplification. NASBA is applicable to more body fluids and requires less plasma (100 uL) than the Amplicor test. Its clinical sensitivity is 400 molecules, and the assay is reproducible and interpretable over a four-log range. Quantiplex HIV RNA assay (bDNA) is a signal, rather than target, amplification method, allowing direct measurement of HIV RNA from 1 mL plasma specimens. Detection (via chemoluminescent substrate) requires hybridization of probes and branch DNA (bDNA) amplifier molecules to HIV RNA. The assay's dynamic range is currently 10,000 - 1,600,000 HIV RNA equivalents/mL, and it is about 20-fold less sensitive than the other two assays. Its detection ability is inversely related to the CD4 cell count.^ieng

[False positive results of HIV virus tests in patients undergoing chronic hemodialysis]. / Anti-human immundeficiencia vírus (HIV) álpozitivitás vizsgálata krónikus haemodialysissel kezelt betegekben.

The sera of 173 haemodialysis patients treated in two dialysis centers in Hungary were tested for the presence of HIV (HTLV III/LAV) antibodies. Four different commercial enzyme immunoassay (EIA) kits and two types (CEM/LAV, and H9/HTLV III) of indirect immunofluorescence assay (IFA) were used. The Western blot technique was applied as confirmatory test in the study. No confirmed positive results were found in any of the cases. However, in 15 patients (8.7%) false positive (not confirmable by the Western blot assay) results were obtained in at least one but mostly in all of the three type 1 EIA kits (ORGANON, ELECTRONUCLEONICS, SORIN) applied. In 4 patients, the IFA assay also gave false positive results which could be repeated in sequential samples taken from the same patients. Increased reactivity in the control plate (coated with a concentrate of cellular material shed by uninfected H9 cell line) of the SORIN kit was found only in a few false positive samples and no fluorescence with the uninfected H9 or CEM cells was observed in any of the sera showing a false positive IFA. These results indicate that the false positive anti-HIV results frequently observable in haemodialysis patients are not simply the consequence of the presence of antibodies reacting with the uninfected H9 and/or CEM cells but they are most probably due to antibodies against antigens expressed on these cells only after infection with the human immunodeficiency virus.

[Analysis of specificity in the serodiagnosis of acquired immunodeficiency syndrome]. / Analiz spetsifichnosti serodiagnostiki sindroma priobretennogo immunnogo defitsita.

Analysis of serum samples for the presence of antibody to AIDS-inducing virus was carried out with the use of the Virognostika diagnosticum manufactured by Organon (the Netherlands). As a result, 6.1% of sera from patients with hematologic diseases, 5.7% of sera from oncologic patients, and 9.2% of sera from hepatitis B patients proved to be "positive" according to the criteria of the manufacturer. However the positively reacting sera could be neutralized by sorption on human red cells, groups AB and 0 and by treatment with the lysate of human lymphocyte culture, which was indicative of the fact that false positive reactions were probably due to the contamination of the antigenic preparation by components of the producer-cell membrane.

[Studies of AIDS in Lithuania]. / Issledovaniia SPID v Litve.

The seroepidemiological survey of 400,000 persons aged 20-40 years and belonging to different AIDS risk groups, as well as blood donors, for the presence of antibodies to HIV has been carried out on the territory of Lithuania. This investigation has been made with the use of the assay systems "Antigen", "Peptoscreen" and "Vector" manufactured in the USSR, as well as commercial assay systems from foreign manufacturers, such as Du Pont de Nemours Inc., Organon N. V., Abbott Laboratories, Serodia. The comparison of the results thus obtained has revealed that high frequency of false positive results is characteristic of all assay systems under study, including immunoblotting. These data indicate that test systems based on different acting principles should be used for the detection of anti-HIV antibodies. For the first time a HIV-infected resident of Lithuania has been detected. The investigation carried out in Lithuania has shown that HIV infection is not widely spread in this region, but due to some objective reasons this does not preclude the necessity of the constant epidemiological surveillance of this infection throughout this territory in order to bar the way to this infection.

[Comparative evaluation of BACTEC MGIT 960 system with MB/BacT and egg-based media for recovery of mycobacteria].

Fully automated, nonradiometric mycobacteria culture systems, BACTEC MGIT 960 (Becton Dickinson Microbiology Systems, Sparks, Md, U.S.A.) and MB/BacT (Organon Teknika, Durham, NC, U.S.A.) were evaluated in comparison with three different eggbased media (3% Ogawa, Ogawa K, and Vite) for the ability to detect mycobacteria in clinical sputum specimens. Sputum specimens were processed by semi-alkaline protease-N-acetyl-L-cysteine-NaOH (SAP-NALC-NaOH) for the automated systems, and by cetylpyridium chloride-succinic acid-NaCl for the egg-based media. A total of 954 sputum specimens were processed, and the recovery of mycobacteria by the BACTEC MGIT 960 was performed in a commercial laboratory. Overall, the frequency of breakthrough contamination was <1% for the three egg-based media, ranging from 0. 42% to 0.63%. Whereas, the frequency of false positives due to breakthrough contamination was 1.89% for MB/BacT and 20.1% for BACTEC MGIT 960. A total of 237 isolates of Mycobacterium tuberculosis complex and 167 isolates of nontuberculous mycobacteria (NTM) were recovered. The highest recovery ratio was obtained by MB/BacT (95.8%), followed by the egg-based media, Vite (74.3%), Ogawa K (65.8%), and 3% Ogawa (58.9%). The recovery ratio by BACTEC MGIT 960 was the lowest, and estimated at 43.1%, mainly due to a high frequency of breakthrough contamination. However, even after omission of these false positives reported by BACTEC MGIT 960, the recovery ratio by this system was comparable to that of 3% Ogawa media. The time to detection of 50% of positive cultures of M. tuberculosis complex by BACTEC MGIT 960 and MB/BacT was 20 days and 17 days, respectively.>From these results, it may be concluded that MB/BacT is superior to BACTEC MGIT 960 and egg-based media for the recovery of mycobacteria from sputum specimens. Furthermore, based on the outcome of this study, we think that considerable improvements are necessary for the clinical application of BACTEC MGIT 960. These improvements should particularly be focused on reducing the false positive ratio caused by contamination, and culture media, which effectively support mycobacterial growth.

[Evaluation of a fully automated mycobacteria culture system, MB/BacT using a newly developed digestion-decontamination procedure, semi-alkaline protease-N-acetyl-L-cysteine-NaOH (SAP-NALC-NaOH) method].

A fully automated non-radiometric mycobacteria culture system, MB/BacT (Organon Teknika, Durham, NC, U.S.A.), was recently introduced in Japan and evaluated for its ability to detect mycobacteria in clinical sputum specimens. A previous study yielded nearly a 40% contamination ratio from sputa treated with the standard N-acetyl-L-cysteine (NALC)-NaOH method. This study employed a mucolytic agent (semi-alkaline protease; SAP) in which the sputa were processed twice for digestion followed by decontamination at twice the standard volume of NALC-NaOH. The concentrated sediments were resuspended in phosphate buffer (0.067 M, pH 6.8), and inoculated into the MB/BacT Process Bottles supplemented with antibiotics. The bottles were incubated at 37 degrees C and monitored for up to fifty-six days. Recovery of mycobacteria was compared in three different egg-based Ogawa media in addition to a non-selective Middlebrook 7H10 agar. A total of 1, 124 clinical sputum specimens have been evaluated. Of these, 464 were positive for growth of mycobacteria, of which 447 (96.3%) were positive by the MB/BacT. False-positive alarms due to break through contamination, mainly by Pseudomonas aeruginosa and Candida spp., were observed in twenty-one specimens (1.9%). The three Ogawa media could detect only 283 (60.5%) to 353 (75.4%) positives, and Middlebrook 7H10 agar only 424 (90.6%) positives. The time to detect positive cultures of Mycobacterium tuberculosis complex by the MB/BacT ranged from 2.2 days to 52.3 days, and 50% of positive cultures were detected within 16.7 days of incubation. It can be concluded that the combination of SAP-NALC-NaOH digestion-decontamination procedure and the MB/BacT is particularly useful for the isolation of mycobacteria and has a faster time to detect than conventional methods. MB/BacT is a suitable alternative method for the detection of mycobacteria in Japan, where the radiometric Bactec System is not available.

[The serodiagnosis of viral hepatitis C]. / Serodiagnostika virusnogo gepatita C.

Sensitivities and specificities of four commercial enzyme immunoassay test systems manufactured by Ortho (USA), Abbott (USA), Organon (Netherlands), Innotest (Belgium) and of three pilot Russian diagnostic agents created at the Mazai Research and Production Amalgamation (Moscow) and the D. I. Ivanovskii Institute of Virology (Moscow) and Pasteur Institute (St. Petersburg) for detection of antibody to hepatitis C have been compared. A high incidence of hepatitis C virus infection was revealed in various Moscow populations: blood and plasma donors, inpatients, and medical staff.

Effects of potentised substances on growth rate of the water plant Lemna gibba L.

OBJECTIVES: This study investigated, whether the growth rate of Lemna gibba L. (duckweed) can be influenced by the application of homeopathic potencies of gibberellic acid, kinetin, argentum nitricum, and lemna minor. METHODS: Duckweed was grown in either potencies (14x-30x, decimal steps) or water controls (unsuccussed and succussed) over seven days. Frond (leaf-like structure) growth was measured using a non-destructive image analysis system. Growth rates were calculated for three time intervals (0-7, 0-3, 3-7 days). Five to six independent, randomized and blinded experiments were analysed for each of the four tested substances. Water control experiments were performed repeatedly to test the reliability of the experimental set-up (systematic negative controls). RESULTS: The systematic negative control experiments did not yield any significant effects. Hence, false positive results could be excluded. The test system had a low coefficient of variation (1.5%). Out of the four tested substances gibberellic acid had the most pronounced effect (p=0.0002, F-test) on the main outcome parameter frond growth rate (r(area) day 0-7). Potency levels 15x, 17x, 18x, 23x and 24x reduced growth rate of Lemna gibba (p<0.05 against the pooled water control, LSD test). CONCLUSIONS: Lemna gibba may be considered as a suitable test organism for further studies on the efficacy of homeopathic potencies. Evidence accumulates, that adjacent potency levels may strongly differ in their biological activity. Potential consequences for therapeutical application might be worth investigating.

[Comparison of the automatized BacT/ALERT blood culture incubation and reading system and a conventional system]. / Comparación del sistema automatizado de incubación y lectura de hemocultivos BacT/ALERT con un sistema convencional.

BACKGROUND: The aim of the present study was to compare the BacT/ALERT blood culture automatic management and reading system (Organon Teknika) with a conventional, nonautomatic technique (DUO, Bio-Mérieux). METHODS: 1,405 blood cultures were parallel compared. 263 of them were positive; out of these, 148 were considered as indicative of septicaemia, 38 of doubtful clinical significance and 77 as accidental contaminations. RESULTS: No differences were detected between both systems neither in the number of isolates obtained nor in the kind of microorganisms recovered or in the number of false-positive or false-negative readings. The conventional system detected 28% of significant isolates during the first 24 hours, reaching 77% after 48 hours; whereas the BacT/ALERT system had detected 57% of significant isolates after 12 hours, reaching 82% after 24 hours (p < 0.001). CONCLUSIONS: The results obtained and the automatization of the BacT/ALERT system appoint it as a firm candidate to be included into the Clinical Microbiology laboratory routine.

[Comparative study of three technics of inverse passive hemagglutination in the search for a surface antigen of the hepatitis b virus (Ag HBs) (author's transl)]. / Etude comparative de trois techniques d'hémagglutination passive inverse pour la recherche de l'antigène de surface du virus de l'hépatite B (Ag HBs).

The following three different versions of reverse passive hemagglutination (RPHA) were evaluated for the detection of HBs antigen: Auscell I - Abbott, WH HBs - Wellcome and the hepanosticon - Organon technique and results obtained were compared with those obtained by the radio immuno assay (Ausria II of Abbott). In 493 sera studied, up to 16,8% were found positive by RPHA as compared to 17,2% positives by RIA. The percentage of false positives by the different methods varied from 4,9 to 7,3. Confirmatory tests, either absorption or neutralization, are necessary to ascertain accuracy of positive results in each of the 3 RPHA methods. The high quality of the Auscell and WH HBs confirmative test allows their sole use although they are slightly less sensitive than the RIA. We would recommand use of the Hepanosticon test whenever positive sera can be confirmed by RIA.

Evaluation of three second-generation and three confirmatory assays for antibodies to human immunodeficiency virus.

The second-generation enzyme immunoassays (EIAs) for antibodies against human immunodeficiency virus (HIV) from three manufacturers (Abbott, Organon, Wellcome) and three anti-HIV confirmatory tests, i.e. Western Blot (WB, Biotech, Dupont), radioimmunoprecipitation assay (RIPA, CLB) and a competitive immunoassay (CIA, Abbott) were evaluated on a panel of 6,488 serum samples, which had previously been used for the comparison of seven first-generation EIAs. For the present study the panel was expanded with sequential serum samples from 12 individuals followed at 1- to 3-month intervals during seroconversion for anti-HIV. The second-generation EIAs and confirmatory tests were significantly more sensitive than the first-generation EIAs as was demonstrated by detection of 10- to 100-fold higher endpoint titers in anti-HIV-positive sera as well as by earlier detection of anti-HIV in 7-11 of the 12 subjects, who seroconverted. In all sera obtained during early HIV infection anti-gp 160/120env antibodies (WB, CIA) were found in addition to anti-p24 (WB, RIPA) and in serial twofold dilutions of these 'seroconversion samples' the new Abbott EIA and RIPA were significantly more sensitive than WB (p less than 0.05), whereas CIA and the new Organon EIA were significantly less sensitive than WB (p less than 0.05). The new Wellcome EIA was not statistically less sensitive than WB. The CIA was as sensitive as WB for antibodies to envelope proteins (gp41, gp160, gp120), but considerably less sensitive for core (p24) antibodies, as was shown in sera obtained during early as well as later clinical stages of HIV infection.(ABSTRACT TRUNCATED AT 250 WORDS)

'Cure' as the gold standard for likelihood ratio assessment: theoretical considerations.

A gold standard is necessary to assess the validity of homeopathic symptoms. The gold standard is 'cure', but this is difficult to define, and depends on consensus. The likelihood ratio (LR) method will give valid results only if the gold standard is reliable. False positives (patients incorrectly classified as cured) weaken results of LR investigation. Weakening the standard to enlarge the research population will seriously bias the results. The same gold standard should be used in LR assessment of all symptoms.

Evaluation of six enzyme immunoassays for antibody against human immunodeficiency virus.

Six commercial enzyme immunoassays (EIA) were evaluated in 6488 serum samples, with immunoblot analysis as the confirmatory test for antibodies against human immunodeficiency virus (HIV). The Abbott and Wellcome tests identified all 163 immunoblot-positive samples correctly, whereas the other tests did not detect 1-3 samples. In AIDS patients (predominantly with antibodies to gp41env) Organon's EIA was less sensitive (p less than 0.05) and Wellcome's more sensitive (p less than 0.05) than the immunoblot assay. In symptom-free anti-HIV-positive subjects (antibodies to almost all viral antigens and high titres of anti-p24gag) all the EIA were significantly (p less than 0.05) less sensitive than the immunoblot assay. The frequencies of false-positive reactions in a "tricky" panel of samples from patients with autoimmune and acute viral diseases and in a blood-donor panel were Abbott 9.5%, 0.42%: Organon 1.7%, 0%; Litton 1.0%, 0.4%; Behring 2.7%, 0.06%; Wellcome 0%, 0%; and Pasteur 0%, 0.02%. The results of a seventh EIA (Dupont) were excluded from the study at the company's request. All six EIA evaluated are suitable tests for anti-HIV screening in samples from patients and blood donors.

Discordant results in human chorionic gonadotropin assays.

Discordance has been reported in human chorionic gonadotropin (hCG) concentrations measured by different immunoassay kits. We examined the results for 40 serum samples assayed with 10 different hCG immunoassay kits. Results varied considerably. Individual sample results varied by as much as 58-fold. Average results for different kits varied by as much as 1.4-fold for pregnancy (20 samples) and 2.2-fold for trophoblast disease (20 samples) serum. We investigated the causes of this discordance. hCG or hCG beta are general names for mixtures of hCG, hCG alpha, or hCG beta immunoreactive molecules in serum. These mixtures include regular hCG, nicked hCG (missing peptide linkages at beta 44-45 or beta 47-48), carbohydrate variants of hCG, hCG missing the beta-subunit C-terminal segment, free beta-subunit, beta-core fragment, and free alpha-subunit. We prepared standards for each of these major variants and measured their reactivities in the 10 hCG immunoassay kits. Free beta-subunit reactivity varied from nonrecognition (anti-beta:anti-alpha type kits; Hybritech Tandem-R and others) to overrecognition (one kit had five-fold greater affinity for free beta than for hCG). Kits with antibodies to beta-subunit C-terminal segment (Organon NML and others) failed to recognize hCG missing this segment, a component of serum hCG in trophoblast disease. Kits with anti-hCG antibodies (Serono MAIA-clone and others) had minimal recognition of nicked hCG (12%), a component of all serum hCG samples, and consistently gave the lowest values with all serum samples. We conclude that differences in recognition of nicked hCG, free beta, and these other hCG variants cause discordance in hCG immunoassay results.