Improved Diagnosis of Tuberculous Meningitis Using a Combination of Multiplex Antigens and Antibodies Testing Methods

Introduction: Various serological assays exists, including Antigen and Antibody detection (IgM and IgG) for diagnosis of Tuberculous Meningitis (TBM) cases. To the best of our knowledge, most of the laboratories either do Antigen detection or IgM or IgG, at a time. As different antigens get expressed in a different stage of infection it may be possible that many cases remain undiagnosed due to one test at a time approach. Aim: To evaluate the combination of Mycobacterium Tuberculosis (MTB) antigen (Ag85 Complex and Rv2623) and antibody (Anti-Ag85, Anti-45kD, Anti-HSP-16, Anti-CFP-10 Anti-GroES and Anti-ESAT-6) immunoassay panels in the Cerebrospinal Fluid (CSF) samples for diagnosis of TBM patient. Materials and Methods: In the present prospective study conducted at Central India Institute of Medical Sciences (CIIMS) from October 2013 to April 2015, a total of 200 CSF samples of different groups {confirmed TBM (n=100) and noninfectious neurological diseases as control (n=100)} were analysed by Enzyme-Linked Immunosorbent Assay (ELISA). A panel of MTB antigens consisting of Ag85B, 45kDa, HSP-16, CFP-10, GroES and ESAT-6 were used for detection of antibodies response, whereas polyclonal antibodies were used for antigen detection of Ag85 complex and Rv2623 in the CSF samples. The comparison of the CSF parameter between TBM and non-TBM patients was performed using a student t-test. A p-value <0.05 was considered statistically significant for all the analyses. Results: The study population has similar age and sex distribution (p>0.05). Symptoms of headache, fever, neck stiffness, vomiting, abnormal behaviour, unconsciousness were more common among the TBM patients as compared to non-TBM patients (p<0.05) (TBM Vs non-TBM). Similarly TBM patients had an increase (p<0.05) Vs non-TBM total cell count, protein, parallel blood sugar and decline in CSF sugar and Parallel blood sugar ratio (p<0.05) (TBM Vs non-TBM). We found diagnostic accuracy of 67% to 76% with either antigen or antibody assay, however, combinations of antigen and antibody immunoassay together increase the diagnostic accuracy of up to 96%. Conclusion: Our study recommends that a combination of antigen and antibody assay should be considered for early and accurate diagnosis of TBM cases.

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Serodiagnosis of parasitic diseases.

Clinical Microbiology Reviews ◽

10.1128/cmr.4.4.457 ◽

1991 ◽

Vol 4 (4) ◽

pp. 457-469 ◽

Cited By ~ 24

Author(s):

S E Maddison

Keyword(s):

Monoclonal Antibodies ◽

Field Studies ◽

Antigen Detection ◽

Skin Tests ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Parasitic Diseases ◽

Patient Response ◽

Biological Technology ◽

Antibody Assays

In this review on serodiagnosis of parasitic diseases, antibody detection, antigen detection, use of monoclonal antibodies in parasitic serodiagnosis, molecular biological technology, and skin tests are discussed. The focus at the Centers for Disease Control on developing improved antigens, a truly quantitative FAST-enzyme-linked immunosorbent assay, and the very specific immunoblot assays for antibody detection is highlighted. The last two assays are suitable for field studies. Identification of patient response in terms of immunoglobulin class or immunoglobulin G subclass isotypes or both is discussed. Immunoglobulin isotypes may asist in defining the stage of some diseases. In other instances, use of a particular anti-isotype conjugate may increase the specificity of the assay. Monoclonal antibodies have played important roles in antigen purification and identification, in competitive antibody assays with increased sensitivity and specificity, and in assays for antigen detection in serum, body fluids, or excreta. Molecular biological technology has allowed significant advances in the production of defined parasitic serodiagnostic antigens.

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Detection of Mycobacterial Lipoarabinomannan with A Monoclonal Antibody Qualitative ELISA in Urine of Tuberculous Meningitis Patients

The Indonesian Biomedical Journal ◽

10.18585/inabj.v8i1.9 ◽

2016 ◽

Vol 8 (1) ◽

pp. 55

Author(s):

Sylvia Rachmayati ◽

Anita Liliana Susanti ◽

Basti Andriyoko

Keyword(s):

Monoclonal Antibody ◽

Urine Sample ◽

Tuberculous Meningitis ◽

Diagnostic Tool ◽

Clinical Laboratory ◽

Antigen Detection ◽

Diagnostic Approach ◽

Enzyme Linked Immunosorbent Assay ◽

Mycobacterial Antigen ◽

Radiological Criteria

BACKGROUND: Tuberculous meningitis is the most severe manifestation of tuberculosis. The diagnostic approach of tuberculous meningitis is difficult. Combination of clinical, laboratory and radiological criteria were used in diagnostic approach of tuberculous meningitis. Urinary mycobacterial lipoarabinomannan (LAM) antigen detection is a promising diagnostic tool. Detection of mycobacterial antigen in concentrated urine sample is predicted to improve the positivity rate of the qualitative enzyme-linked immunosorbent assay (ELISA) diagnostic tool. The purpose of this study is to examine the detection ability of a monoclonal antibody qualitative ELISA in concentrated and unconcentrated urine of tuberculous meningitis patients.METHODS: This research is a descriptive, crosssectionally designed. The study was conducted in the Clinical Pathology Department laboratory of Dr. Hasan Sadikin Hospital, in July-October 2014. A total of 27 patients diagnosed as tuberculous meningitis patients were included and the subjects were classified into possible and probable criteria according to consensus criteria. The subjects were classified as definite if the cerebrospinal fluid culture was positive for Mycobacterial tuberculosis growth. The subjects were examined for the presence of LAM in unconcentrated and concentrated urine with a monoclonal antibody qualitative ELISA method.RESULTS: Unconcentrated urinary LAM examination positivity was 0% while in concentrated urine was 14.8%. The positivity of concentrated urinary LAM were higher among the definite criteria group.CONCLUSION: Concentrating urine sample increase the positivity rate of urinary LAM detection with ELISA method as high as 14.8%. The urinary antigen detection is higher among the definite tuberculous meningitis patients.KEYWORDS: LAM, concentrated urine, tuberculous meningitis, qualitative ELISA

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Evaluation of purified 27.5 kDa protoscolex antigen-based ELISA for the detection of circulating antigens and antibodies in sheep and human hydatidosis

Journal of Helminthology ◽

10.1017/s0022149x14000479 ◽

2014 ◽

Vol 89 (5) ◽

pp. 577-583 ◽

Cited By ~ 8

Author(s):

I.R. Bauomi ◽

A.M. El-Amir ◽

A.M. Fahmy ◽

R.S. Zalat ◽

T.M. Diab

Keyword(s):

Recombinant Antigen ◽

Antigen Detection ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Hydatid Cysts ◽

Elisa Method ◽

Active Infection ◽

Diagnostic Efficacy ◽

Detection Assay ◽

Circulating Antigens

AbstractHydatidosis is a zoonotic disease caused by the larval stage of Echinococcus granulosus, and the diagnosis of hydatidosis to date remains unresolved despite the development of many serological techniques. The present study aimed to develop an antigen-based enzyme-linked immunosorbent assay (ELISA) using IgG anti-27.5 kDa protoscolex antigen (27.5 PA) for measuring circulating protoscolex antigen (CPA), for comparison with an antibody detection assay, in sera of naturally infected sheep and humans in highly endemic areas in Egypt. In sheep, the sensitivity of ELISA in detecting anti-27.5 PA IgG and CPA was 75.0 and 60.0%, respectively, and the recorded specificity was 80.0 and 88.0%, respectively. In humans, the sensitivity of ELISA in detecting anti-27.5 PA IgG and CPA was 62.5 and 52.5%, respectively, while the specificity of the assay was 66.7 and 75.0%, respectively. In conclusion, an antibody detection assay is still superior and is more sensitive than an antigen detection assay, especially in diagnosing an active infection in which hydatid cysts are predominant. An antigen detection assay may be a useful approach to assessment of the efficacy of treatment, especially after removal of the cyst. Further studies are recommended to improve the diagnostic efficacy of an antigen-based ELISA method by using a highly purified recombinant antigen.

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Diagnostic accuracy of two commercially available rapid assays for detection of IgG and IgM antibodies to SARS-CoV-2 compared to ELISA in a low-prevalence population

10.21203/rs.3.rs-50887/v1 ◽

2020 ◽

Author(s):

Klaus Hackner ◽

Peter Errhalt ◽

Martin Willheim ◽

Maria-Anna Grasl ◽

Jasmina Lagumdzija ◽

...

Keyword(s):

Diagnostic Accuracy ◽

Close Contact ◽

Point Of Care ◽

Rapid Test ◽

Screen Test ◽

Elisa Test ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Rapid Assays ◽

Low Prevalence

Abstract Background: New commercially available point-of-care (POC) immunodiagnostic tests are appearing, which may yield rapid results for anti-SARS-CoV-2 antibodies. The aim of this study was to evaluate the diagnostic accuracy of rapid antibody detection tests compared to a validated laboratory-based enzyme-linked immunosorbent assay (ELISA) and to investigate infections amongst healthcare workers (HCWs) after unprotected close contact to COVID-19 patients. Methods: Blood serum and whole blood of 130 participants were tested with NADAL® COVID-19 IgG/IgM Rapid Test and mö-screen 2019-NCOV Corona Virus Test against a validated ELISA test. Infection status was evaluated using real-time polymerase-chain-reaction.Results: Acute COVID-19 infection was detected in 2.4% of exposed HCWs. Antibody tests showed an overall frequency of IgG and IgM in 5.3%, with 1.6% asymptomatic infections. The NADAL® test showed a sensitivity (IgM/ IgG) of 100% (100%/ 100%), a specificity (IgM/ IgG) of 98.8% (97.6%/ 100 %), a PPV of 76.9% (57.1%/ 100%), an NPV of 100% (100%/ 100%), and a diagnostic accuracy of 98.8% (97.7%/ 100%). The mö-screen test had a sensitivity (IgM/IgG) of 90.9% (80%/ 100%), a specificity (IgM/IgG) of 98.8% (97.6%/ 100%), a PPV of 76.9% (57.1%/ 100%), an NPV of 99.6% (99.2%/ 100%), and a diagnostic accuracy of 98.5% (96.9%/ 100%). Conclusions: The frequency of COVID-19 infections in HCWs after unprotected close contact is higher than in the general population of a low-prevalence country. Both POC tests were useful for detecting IgG, but did not perform well for IgM, mainly due to false positive results.

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Detection of Humoral Antigen and Antibody by Enzyme-Linked Immunosorbent Assay in Horses with Experimentally Induced Ehulichia Equi Infection

Journal of Veterinary Diagnostic Investigation ◽

10.1177/104063879300500109 ◽

1993 ◽

Vol 5 (1) ◽

pp. 37-39 ◽

Cited By ~ 14

Author(s):

Richard E. Corstvet ◽

Stephen D. Gaunt ◽

Phillip A. Karns ◽

Jere W. McBride ◽

Ricardo A. Battistini ◽

...

Keyword(s):

Monoclonal Antibodies ◽

Immunosorbent Assay ◽

Rectal Temperature ◽

Diagnostic Method ◽

Serum Antibody ◽

Clinical Signs ◽

Antigen Detection ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Ehrlichia Equi

An enzyme-linked immunosorbent assay (ELISA) was used to detect antigen in plasma and antibody in serum of 3 horses inoculated with Ehrlichia equi. Clinical signs, including rectal temperature, were correlated with the antigen and antibody detection. ELISA was very efficient in detection of serum antibody. Antigen detection using monoclonal antibodies to E. equi and ELISA should be considered as a diagnostic method.

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POS0441 DEVELOPMENT OF RHEUMATOID ARTHRITIS AMONG ANTI-CITRULLINATED PROTEIN ANTIBODIES POSITIVE ASYMPTOMATIC INDIVIDUALS: A PROSPECTIVE OBSERVATIONAL STUDY

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-eular.344 ◽

2021 ◽

Vol 80 (Suppl 1) ◽

pp. 449.1-449

Author(s):

S. Mizuki ◽

K. Horie ◽

K. Imabayashi ◽

K. Mishima ◽

K. Oryoji

Keyword(s):

Rheumatoid Arthritis ◽

Risk Factors ◽

Observational Study ◽

Rheumatoid Factor ◽

Prospective Observational Study ◽

Enzyme Linked Immunosorbent Assay ◽

P Value ◽

Healthy Population ◽

Asymptomatic Individuals ◽

Citrullinated Protein

Background:In the idividuals with genetic and enviromental risk factors, immune events at mucosal surfaces occur and may precede systemic autoimmunity. Anti-citrullinated protein antibodies (ACPA) are present in the serum for an average of 3-5 years prior to the onset of rheumatoid arthritis (RA) during an asymptomatic period. In ACPA-positivite individuals, the additional presence of RA-related risk factors appears to add significant power for the development of RA. To date, there have been few reports in which clinical courses of ACPA-positive asymptomatic individuals were investigated prospectively.Objectives:To observe the clinical time course of ACPA-positive healthy population for the development of RA.Methods:Healthy volunteers without joint pain or stiffness, who attended the comprehensive health screening of our hospital, were enrolled in this prospective observational study. The serum ACPA levels were quantified by Ig-G anti-cyclic citrullinated peptide enzyme-linked immunosorbent assay with levels > 4.4 U/mL considered positive. ACPA-positive subjects were followed by rheumatologists of our department clinically or a questionnaire sent by mail for screening to detect arthritis.Results:5,971 healthy individuals without joint symptons were included. Ninty-two (1.5%) were positive for ACPA. Of these, 19 (20.7%) developed RA and two were suspected as RA by mail questionnaire. Their average age were 58-years, and women were 68%. The average duration between the date of serum sampling and diagnosis was 10.7 months. ACPA-positive individuals who developed to RA had higher serum ACPA and Ig-M rheumatoid factor levels than ACPA-positive individuals who did not (P value by Mann-Whitney U test: 0.002, 0.005, respectively).Conclusion:Among ACPA-positive asymptomatic individuals, 20% developed RA. The higher titer of ACPA and Ig-M rheumatoid factor levels are risk factors for devoloping RA.Disclosure of Interests:None declared

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Validation of a Novel Commercial ELISA Test for the Detection of Antibodies against Coxiella burnetii

Pathogens ◽

10.3390/pathogens9121075 ◽

2020 ◽

Vol 9 (12) ◽

pp. 1075

Author(s):

Salvatore Ledda ◽

Cinzia Santucciu ◽

Valentina Chisu ◽

Giovanna Masala

Keyword(s):

Diagnostic Accuracy ◽

Phase Ii ◽

Coxiella Burnetii ◽

Q Fever ◽

Animal Health ◽

Elisa Test ◽

Clinical Diagnostics ◽

Complex Life Cycle ◽

Enzyme Linked Immunosorbent Assay ◽

Specificity And Sensitivity

Q fever is a zoonosis caused by Coxiella burnetii, a Gram-negative pathogen with a complex life cycle and a high impact on public and animal health all over the world. The symptoms are indistinguishable from those belonging to other diseases, and the disease could be symptomless. For these reasons, reliable laboratory tests are essential for an accurate diagnosis. The aim of this study was to validate a novel enzyme-linked immunosorbent assay (ELISA) test, named the Chorus Q Fever Phase II IgG and IgM Kit (DIESSE, Diagnostica Senese S.p.A), which is performed by an instrument named Chorus, a new device in medical diagnostics. This diagnostic test is employed for the detection of antibodies against C. burnetii Phase II antigens in acute disease. Our validation protocol was performed according to the Italian Accreditation Body (ACCREDIA) (Regulation UNI CEI EN ISO/IEC 17025:2018 and 17043:2010), OIE (World Organization for Animal Health), and Statement for Reporting Studies of Diagnostic Accuracy (STARD). Operator performance was evaluated along with the analytical specificity and sensitivity (ASp and ASe) and diagnostic accuracy of the kit, with parameters such as diagnostic specificity and sensitivity (DSp and DSe) and positive and negative predictive values (PPV and NPV), in addition to the repeatability. According to the evaluated parameters, the diagnostic ELISA test was shown to be suitable for validation and commercialization as a screening method in human sera and a valid support for clinical diagnostics.

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Exploratory field study on the effects of porcine circovirus 2 (PCV-2) sow vaccination at different physiological stages mimicking blanket vaccination

Porcine Health Management ◽

10.1186/s40813-021-00213-2 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Patricia Pleguezuelos ◽

Marina Sibila ◽

Raúl Cuadrado ◽

Rosa López-Jiménez ◽

Diego Pérez ◽

...

Keyword(s):

Quantitative Polymerase Chain Reaction ◽

Negative Control Group ◽

Negative Control ◽

Late Gestation ◽

Antibody Detection ◽

Porcine Circovirus ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

Physiological Status ◽

Porcine Circovirus 2

Abstract Background The objective of the present study was to explore the benefits of Porcine circovirus 2 (PCV-2) blanket vaccination in a sow herd on productive parameters, PCV-2 infection and immune status in sows and their progeny. For this purpose, 288 sows were distributed among four balanced experimental groups. One group remained as negative control group and the other three received 1 mL of PCV-2 Ingelvac Circoflex® intramuscularly at different productive cycle moments: before mating, mid gestation (42–49 days post-insemination) or late gestation (86–93 days post-insemination); phosphate buffered saline (PBS) was used as negative control item. Reproductive parameters from sows during gestation and body weight of their progeny from birth to weaning were recorded. Additionally, blood was collected from sows at each vaccination time and piglets at 3 weeks of age. Moreover, up to 4 placental umbilical cords (PUC) per sow were taken at peri-partum. Sera from sows and piglets were analysed for PCV-2 antibody detection using an enzyme-linked immunosorbent assay (ELISA). Sera from sows and PUC were tested to quantify viraemia using a real time quantitative polymerase chain reaction (qPCR) assay. Results Globally, results indicated that vaccinated sows showed heavier piglets at birth and at weaning, less cross-fostered piglets, lower viral load at farrowing as well as in PUC, and higher antibody levels at farrowing, compared to non-vaccinated ones. When all groups were compared among them, sows vaccinated at mid or late gestation had heavier piglets at birth than non-vaccinated sows, and lower proportion of PCV-2 positive PUC. Also, cross-fostering was less frequently practiced in sows vaccinated at pre-mating or mid gestation compared to non-vaccinated ones. Conclusions In conclusion, the present study points out that PCV-2 sow vaccination at different time points of their physiological status (mimicking blanket vaccination) offers benefits at production and serological and virological levels.

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Development and evaluation of a monoclonal antibody-based competitive ELISA for the detection of antibodies against H7 avian influenza virus

BMC Veterinary Research ◽

10.1186/s12917-021-02772-6 ◽

2021 ◽

Vol 17 (1) ◽

Author(s):

Yuan Li ◽

Hongliu Ye ◽

Meng Liu ◽

Suquan Song ◽

Jin Chen ◽

...

Keyword(s):

Monoclonal Antibody ◽

Influenza Virus ◽

Avian Influenza ◽

Avian Influenza Virus ◽

Large Scale ◽

Operation Time ◽

Reference Method ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Antibody Concentration

Abstract Background H7 subtype avian influenza has caused great concern in the global poultry industry and public health. The conventional serological subtype-specific diagnostics is implemented by hemagglutination inhibition (HI) assay despite lengthy operation time. In this study, an efficient, rapid and high-throughput competitive enzyme-linked immunosorbent assay (cELISA) was developed for detection of antibodies against H7 avian influenza virus (AIV) based on a novel monoclonal antibody specific to the hemagglutinin (HA) protein of H7 AIV. Results The reaction parameters including antigen coating concentration, monoclonal antibody concentration and serum dilution ratio were optimized for H7 antibody detection. The specificity of the cELISA was tested using antisera against H1 ~ H9, H11 ~ H14 AIVs and other avian viruses. The selected cut-off values of inhibition rates for chicken, duck and peacock sera were 30.11, 26.85 and 45.66% by receiver-operating characteristic (ROC) curve analysis, respectively. With HI test as the reference method, the minimum detection limits for chicken, duck and peacock positive serum reached 20, 21 and 2− 1 HI titer, respectively. Compared to HI test, the diagnostic accuracy reached 100, 98.6, and 99.3% for chicken, duck and peacock by testing a total of 400 clinical serum samples, respectively. Conclusions In summary, the cELISA assay developed in this study provided a reliable, specific, sensitive and species-independent serological technique for rapid detection of H7 antibody, which was applicable for large-scale serological surveillance and vaccination efficacy evaluation programs.

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Prevalence and clinical correlation of hepatitis E virus antibody in the patients’ serum samples from a tertiary care hospital in Thailand during 2015–2018

Virology Journal ◽

10.1186/s12985-021-01616-x ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Atiporn Boonyai ◽

Anchalee Thongput ◽

Thidarat Sisaeng ◽

Parisut Phumchan ◽

Navin Horthongkham ◽

...

Keyword(s):

Pregnant Women ◽

Tertiary Care ◽

Laboratory Data ◽

Organ Transplant ◽

Hospital Setting ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Care Hospital ◽

Serum Samples ◽

Igm Antibodies

Abstract Background Prevalence and incidence of hepatitis caused by HEV infection are usually higher in developing countries. This study demonstrated the HEV seroprevalence and incidence of HEV infection in patients with clinical hepatitis in a tertiary hospital in Thailand. Methods A laboratory-based cross-sectional study was conducted using 1106 serum samples from patients suspected of HEV infection sent to the Serology laboratory, Siriraj Hospital, for detecting HEV antibodies during 2015–2018. Prevalence of anti-HEV IgG and IgM antibodies in general patients, including organ transplant recipients and pregnant women in a hospital setting, were determined using indirect enzyme-linked immunosorbent assay (ELISA) kits. Comparison of laboratory data between groups with different HEV serological statuses was performed. Results HEV IgG antibodies were detected in 40.82% of 904 serum samples, while HEV IgM antibodies were detected in 11.75% of 1081 serum samples. Similar IgG and IgM antibody detection rates were found in pregnant women. Interestingly, anti-HEV IgM antibodies were detected in 38.5% of patients who underwent organ transplantation. Patients who tested positive for anti-HEV IgM antibodies had higher alanine aminotransferase levels than those who had not. In contrast, patients who tested positive for anti-HEV IgG had more elevated levels of total bilirubin than those who tested negative. Conclusions HEV seroprevalence and incidence in patients with clinical hepatitis were relatively high in the Thai population, including the pregnancy and organ transplant subgroups. The results potentially benefit the clinicians in decision-making to investigate HEV antibodies and facilitating proper management for patients.

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