• Title/Summary/Keyword: false-positive reaction

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Clearance of False-positive Antigen-Antibody Reactions of a Diagnostic Antigen Production in Escherichia coli with Human Sera

  • Noh, Kap-Soo;Kim, Jong-Wan;Ha, Suk-Hoon;Yoo, Wang-Don;Jeon, Weong-Joong;Kim, Hyun-Su
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.4 no.1
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    • pp.63-65
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    • 1999
  • Although many pharmaceutically useful proteins are produced in E. coli expression system, it is very are rare for the system to be used in the production of diagnostic antigen due to a major problem, i.e., false-positive reaction of e. coli host-derived proteins contaminating purified diagnostic antigen with human sera. The N (nucleocapsid) protein of Seoul virus causing haemorrhagic fever with renal syndrome (HFRS) was produced in E. coli BL21 (DE3), and used for the detection of N protein-specific antibodies in human sera. Using the N protein as a diagnostic antigen of HFRS, the false-positive reaction was cleared by merely mixing the test sera with the extract of E. coli host strain not harboring expression plasmid.

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Effect of ethylenediamine on luminol (or Bluestar) - bleach reaction (Ethylenediamine이 luminol (or Bluestar) - 표백제 반응에 미치는 영향)

  • Seulgi, Jang;Minkyoung, Kim;Heejin, Kim;Munhee, Lee;Sungwook, Hong
    • Analytical Science and Technology
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    • v.35 no.6
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    • pp.242-248
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    • 2022
  • The effect of ethylenediamine (EDA) on the reaction of luminol or Bluestar with blood and bleaches was studied. For this purpose, blood, chlorine bleach, and oxygen bleach were applied to filter paper, treated with EDA-containing luminol or Bluestar, and the changes in chemiluminescence intensity were observed. It was found that the chemiluminescence intensity of the luminol (or Bluestar)-blood reaction did not change with the increasing concentration of EDA. However, the chemiluminescence intensity of the luminol (or Bluestar)-chlorine bleach reaction decreased and the chemiluminescence intensity of the luminol (or Bluestar)-oxygen bleach reaction increased, with increasing EDA concentration. Thus, it was found that when EDA was added to luminol (or Bluestar), which is a blood-sensitive reagent, EDA suppressed the false-positive reaction induced by chlorine bleach and induced a false-positive reaction with oxygen bleach. Consequently, the addition of EDA is not recommended for enhancing bloodstains with luminol or Bluestar.

A Simple Method for Elimination of False Positive Results in RT-PCR

  • Martel, Fatima;Grundemann, Dirk;Schomig, Edgar
    • BMB Reports
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    • v.35 no.2
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    • pp.248-250
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    • 2002
  • Discrimination between the amplification of mRNA and contaminating genomic DNA is a common problem when performing a reverse transcriptase-polymerase chain reaction (RT-PCR). Even after treatment of the samples with DNAse, it is possible that negative controls (samples in which no reverse transcriptase was added) will give positive results. This indicates that there was amplification of DNA, which was not generated during the reverse transcriptase step. The possibility exists that Taq DNA polymerase acts as a reverse transcriptase, generating cDNA from RNA during the PCR step. In order to test this hypothesis, we incubated samples with a DNAse-free RNAse after the cDNA synthesis. Comparison of the results that were obtained from these samples (incubated with or without DNAse-free RNAse) confirms that the reverse transcriptase activity of Taq DNA polymerase I is a possible source of false positive results when performing RT-PCR from intronless genes. Moreover, we describe here a simple and rapid method to overcome the false positive results that originate by this activity of Taq polymerase.

A study on the false-positive reaction of Kastle-Meyer(KM) test and the application of KM reagent (Kastle-Meyer(KM) test의 위양성 반응과 시약 적용 방법에 관한 연구)

  • Kim, Chae-Lin;Kim, Chang-Yong;Kim, Min-Yeong;Yu, Je-Seol
    • Journal of the Korea Convergence Society
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    • v.11 no.10
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    • pp.139-146
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    • 2020
  • Kastle-Meyer (KM) test is one of presumptive tests used to detect latent blood in crime scenes. While this method is more sensitive than others, false positive reaction can be shown where blood does not exist. In this study, we tried to suggest a new application method to solve this problem. Reaction time and aspect of reaction of this new method was compared with two conventional methods in order to identify the effectiveness and identifiability. As a result, there was no statistically significant difference in sensitivity between the methods. In addition, in case of blood with a dilution ratio of 5000:1 or less, positive reaction showed immediately, making it easy to distinguish the reaction from false positive reaction. However, it became difficult to distinguish them with the reaction time as the dilution ratio increased, and this phenomenon could be supplemented observing aspect of the reaction when using the new method. Therefore, this study suggested a new method for KM test that can be used more accurately in the field.

Relationship Between Skin Impedance Signal, Reaction time, and Eye Blink Depending on Arousal Level (각성상태에 따른 피부임피던스 신호와 반응시간 및 눈 잡학임의 상관관계(E))

  • 고한우;김연호
    • Journal of Biomedical Engineering Research
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    • v.18 no.4
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    • pp.485-491
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    • 1997
  • This paper describes the relationship between skin impedance signal, behavioral signal, and subjective evaluation depending on arousal level. Nz and reaction time had similar trend with mKSS level, but eyeblink rate was different from these two parameters. eye-blink rate increased slowly from mKSS level 1 to 5, and had high increasing rate at mKSS 7. But it showed steep descent at mKSS level 9. Each subject showed different eye-blink rates, but changing rates of EBR was similar at eachm KSS level. Therefore it suggests that rising rate of EBR can be used arousal level criterion. From the result of reaction time test. human performance was decreased rapidly above the mKSS level 5, and false positive and false negative data was observed above the mKSS level 3. It is desirable to give a subject some stimuli such as sound or aroma to rise arousal level between mKSS level 3 and mKSS level 5.

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Protease antigen recovery induces non-specific reaction in B-Cells (Protease antigen recovery의 B-Cell에 대한 비특이반응 유발)

  • Kim, Ok-Jin;Yi, Seong-Joon
    • Korean Journal of Veterinary Pathology
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    • v.7 no.1
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    • pp.11-15
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    • 2003
  • Antigen retrieval (AR) techniques were widely used to recover the antigenicity from the fixed tissues, which were guided by the philosophy of rendering immunohistochemistry (IHC) applicable to routine formalin-fixed, paraffin-embedded tissues for wide application of IHC in research and clinical filed for morphological observation like as anatomy, histology and pathology. Protease antigen recovery (PAR) is an AR technique, which is obtained the antigen retrieve by using enzyme digestion, and commonly used in IHC field. However, during the IHC for the detection of ovine herpesvirus 2 (OvHV-2) antigen, we noted lymphocyte-like cells-specific staining in the infiltrated cells into various organs like as liver and kidney, which was also shown in the IHC tissues with isotype control. However, those signals were not observed in the tissues conducted with in situ hybridization. Therefore, we analyzed the specificity of the IHC detection results. We found that PAR may induce false-positive result during IHC in lymphocyte-like cells, which were infiltrated mainly around vessels and in interstitial tissues. Through the Phenotyping, we realized that those false-positive cells were B-cell-related cells. These results suggest that PAR, a AR using protease, may induce non-specific false-positive reactions during IHC.

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Diagnosis of viral fish diseases by polymerase chain reaction - restriction fragment length polymorphism (Polymerase chain reaction - restriction fragment length polymorphism을 이용한 바이러스성 어류 질병 진단)

  • Kim, Myoung-Sug;Park, Shin-Hoo;Cho, Mi-Young;Kim, Jin-Woo;Park, Myoung-Ae
    • Journal of fish pathology
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    • v.21 no.3
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    • pp.181-188
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    • 2008
  • Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was used to detect and identify four fish viruses, fish iridovirus, viral hemorrhagic septicaemia virus (VHSV), viral nervous necrosis virus (VNNV), hirame rhabdovirus (HRV). Four viruses were detected by PCR with each specific primers. Identification of iridovirus was achieved by digesting the PCR amplified fragment with a restriction enzyme ApaⅠ. It was possible to distinguish positive from false positive PCR amplicons of VHSV by RFLP of PstⅠ or HindⅢ restriction enzymes. VNNV was identified using RFLP of BamHⅠrestriction enzyme and HRV was identified by XbaⅠ restriction enzyme. This approach can be used for more rapid, simple and specific diagnosis of fish viral diseases.

Comparison of RPR Card and Mediace RPR test by KFDA Guideline

  • Lee, Hae Soon
    • Korean Journal of Clinical Laboratory Science
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    • v.44 no.3
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    • pp.124-127
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    • 2012
  • Syphilis is an infectious and sexually transmitted chronic disease caused by Treponema pallidum. On the basis of clinical findings, the disease has been divided into a series of overlapping stages, which are used to help guide treatment and follow-up. Persons who have syphilis might seek treatment for signs or symptoms of primary infection, secondary infection and tertiary infection. Latent infections are detected by serologic testing. A presumptive diagnosis of syphilis is possible with the use of two types of serologic tests: nontreponemal tests and treponemal tests assay. The use of only one type of serologic test is insufficient for diagnosis, because each type of test has limitations, including the possibility of false-positive test results in persons without syphilis. KFDA published Koreans guideline of Sexually transmitted infections in 2011. Two hundred samples were tested by RPR card test and Mediace RPR test with simultaneously. The agreement between RPR card test and Mediace RPR test was 95%, the discrepant samples was 5%. The characteristics of 10 discrepant samples was RPR card Positive and Mediace RPR negative nine samples, RPR card negative and Mediace RPR positive one sample. The nine samples were confirmed as FTA-ABS by KFDA guideline of syphilis test algorism, all IgM test was Negative, all IgG test was reactive. So, these cases were past or latent syphilis. The one sample was false-positive reaction.

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Development of Reverse Transcriptase Polymerase Chain Reaction Primer Sets and Standard Positive Control Capable of Verifying False Positive for the Detection of Severe acute respiratory syndrome coronavirus 2

  • Cho, Kyu Bong
    • Biomedical Science Letters
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    • v.27 no.4
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    • pp.283-290
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    • 2021
  • Severe acute respiratory syndrome coronavirus (SARS-CoV2) is a major coronavirus that infects humans with human Coronavirus (HuCoV)-229E, HCoV-OC43, HCoV-HKU1, HCoV-NL63, Severe acute respiratory syndrome coronavirus (SARS-CoV) and Middle east respiratory syndrome coronavirus (MERS-CoV). SARS-CoV2 is currently a global pandemic pathogen. In this study, we developed conventional RT-PCR based diagnostic system for the detection of SARS-CoV2 which is relatively inexpensive but has high stability and a wide range of users. Three conventional RT-PCR primer sets capable of forming specific band sizes by targeting the ORF1ab [232 nucleotide (nt)], E (200 nt) and N (288 nt) genes of SARS-CoV2 were developed, respectively, and it were confirmed to be about 10~100 times higher detection sensitivity than the previously reported methods. In addition, a standard positive control that can generate specific amplicons by reacting with the developed RT-PCR primers and verify the false-positiv from contamination of the laboratory was produced. Therefore, the diagnostic system that uses the RT-PCR method is expected to be used to detect SARS-CoV2.

Diagnostic Value of the Intradermal Test for the Infection with Clonorchis sinensis (간흡충증 진단을 위한 피내반응검사의 의의)

  • Kim, Jong-Ho;Yun, Bong-Yung;Lee, Heun-Ju;Lee, Hyun-Woo
    • Journal of Yeungnam Medical Science
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    • v.5 no.2
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    • pp.47-52
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    • 1988
  • The intradermal(ID) test has been widely used in Korea and several reports about the results of the ID test are known. We examined the egg of Clonorchis sinensis(C. s.) by ID test in 443, stool's egg-counting technique in 79 and direct smear(cellophane thick smear technique) in 1304 subjects. The results are as follows.: 1. The positive rate of C. s. was 3.8% out of 1304 persons. 2. The sensitivity of ID test was 82.1% out of 39 persons and, the specificity was 64.6% out of 404 persons. 3. The false positive of ID test 35.4% out of 404 persons and, the false negative was 17.9% out of 39 persons. Intradermal test is a rapid, sensitive and useful supplementary diagnostic tool for the detection of Clonorchiasis infection and must be used as screening test with direct smear of stool but cross reaction with other helminth infections and moderate false reaction are the main disadvantages in its practical application.

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