Immunodiagnosis of childhood pulmonary and extrapulmonary tuberculosis usingMycobacterium tuberculosisES antigen by penicillinase ELISA (original) (raw)
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Serodiagnosis of childhood tuberculosis by ELISA
Indian Journal of Pediatrics, 2005
Objective: Diagnosis of childhood tuberculosis remains an enigma despite many recent technological developments. The present study has been taken up with the aim to assess the diagnostic potential of mycobacterium tuberculosis excretory-secretory ES-31 antigen and affinity purified anti ES-31 antibodies in the serodiagnosis of different spectrum of childhood tuberculosis.Methods: Mycobacterium tuberculosis H37 Ra excretory-secretory antigen (ES-31) and affinity purified goat anti ES-31 antibodies were used in stick penicillinase ELISA for IgG antibody detection and stick Sandwich penicillinase ELISA for detection of circulating free and immune complexed antigen in the sera of 230 children.Results: Analysis of tubercular antibody, circulating free and immune complexed antigen (CIC-Ag) was done in both pulmonary and extrapulmonary form of childhood tuberculosis and overall sensitivity of 81.4% with a specificity of 93% was achieved for detection of antitubercular IgG antibodies. Of the five cases of pulmonary tuberculosis showing absence of IgG antibody, 3 showed the presence of CIC-Ag and one was found positive for both free and CIC-Ag. Similarly out of 8 cases of extrapulmonary childhood tuberculosis missed by IgG detection 5 were found to be positive for CIC-Ag and 1 showed the positive reaction for both free and immune complexed antigens.Conclusion: IgG antibody to excretory-secretory antigen ES-31 is found to be having good specificity with acceptable sensitivity in detecting different forms of childhood tuberculosis. Further detection of circulating free and / or immunecomplexed antigen can be used as an adjunct tool in the diagnosis of childhood tuberculosis.
An Enzyme-linked Immunosorbent Assay (ELISA) was established for the detection of IgG antibody in human cerebrospinal fluid (CSF) from patients with tuberculous meningitis to the 30,000-Da native antigen of Mycobacterium tuberculosis. The test was done on patients with bacteriologically confirmed and autopsy-proven tuberculous meningitis (8 cases), clinically presumptive cases of tuberculous meningitis (100 cases), non-tuberculous neurologic disease (88 cases) and normal patients without neurologic disease (10 cases). The test was positive in 7 of the 8 cases of bacteriologically confirmed tuberculous meningitis (87.5%). ELISA was also positive in 67 of 100 cases of clinically presumptive tuberculous meningitis (67.00%). Overall sensitivity was found to be 68.51%. Specificity of ELISA was found to be 97.95% (96/98). The results show the potential use of ELISA as a rapid diagnostic test for tuberculous meningitis. (Phi J Microbiol Infect Dis 2000; 29(4):156-161)
BMC research notes, 2017
Pediatric tuberculous meningitis is a highly morbid, often fatal disease. Its prompt diagnosis and treatment saves lives, in fact delays in the initiation of therapy have been associated with high mortality rates. This is a case of an Italian child who was diagnosed with tuberculous meningitis after a history of a month of headache, fatigue and weight loss. Cerebrospinal fluid analysis revealed a lymphocytic pleocytosis with predominance and decreased glucose concentration. Microscopy and conventional diagnostic tests to identify Mycobacterium tuberculosis were negative, while a non classical method based on intracellular cytokine flow cytometry response of CD4 cells in cerebral spinal fluid helped us to address the diagnosis, that was subsequently confirmed by a nested polymerase chain reaction amplifying a 123 base pair fragment of the M. tuberculosis DNA. We diagnosed tuberculous meningitis at an early stage through an innovative immunological approach, supported by a nested poly...
Indian journal of …, 1987
Bronchial aspirates in 160 children were examined for tubercle bacilfi and were positive in 55 (34"4%). Tubercular antigen was detected in 56 of 79 children (70"9%). In 60 children in whom AFB or TB antigen was positive in branchial washings antigen was positive in more than 96" 7% of cases, indicating high sensitivity of antigen test. Moreover, antigen is specific for human and bovine mycobacteria.Only 15% of children who had tuberculosis proved by + ve antigen or + ve smear presented with usual characteristic symptoms of tuberculosis. About 75% of the children presented with recurrent or persistent bronchitis or pertussoid syndrome, while in 9% of children there were no significant symptoms. Mediastinal lymphnode involvement is common in children and 80~ of AFB/antigen positive cases had only mediastinal lymphnode disease, while 20 also had parenchymal lesions. For the diagnosis of taberculosis a simple technique such as examination of throat secretions for detection of TB antigen by RIA technique or ELISA has great potential.
Immunological Aspects of Diagnosis and Management of Childhood Tuberculosis
Infection and Drug Resistance, 2021
The diagnosis of tuberculosis (TB) in children is difficult because of the low sensitivity and specificity of traditional microbiology techniques in this age group. Whereas in adults the culture of Mycobacterium tuberculosis (M. tuberculosis), the gold standard test, detects 80% of positive cases, it only detects around 30-40% of cases in children. The new methods based on the immune response to M. tuberculosis infection could be affected by many factors. It is necessary to evaluate the medical record, clinical features, presence of drug-resistant M. tuberculosis strains, comorbidities, and BCG vaccination history for the diagnosis in children. There is no ideal biomarker for all TB cases in children. A new strategy based on personalized diagnosis could be used to evaluate specific molecules produced by the host immune response and make therapeutic decisions in each child, thereby changing standard immunological signatures to personalized signatures in TB. In this way, immune diagnosis, prognosis, and the use of potential immunomodulators as adjunct TB treatments will meet personalized treatment.
Diagnostic Value of Elisa Serological Tests in Childhood Tuberculosis
Journal of Tropical Pediatrics, 2006
Two separate studies (study I and study II) were conducted to evaluate the efficacy of ELISA serological test for the detection of IgG antibodies against specific glycolipid antigen (PGLTb1) and ESAT 6 antigen of Mycobacterium tuberculosis, respectively. These results were compared with bacteriological tests [Ziehl Neelson (ZN) staining for acid-fast bacilli and culture on Lowenstein Jensen (LJ) medium] and polymerase chain reaction (PCR) targeting IS6110 sequence. Both studies were carried out on children with pulmonary, central nervous system, lymph node, and gastrointestinal tuberculosis along with matching controls (65 cases and 27 controls for study I and 83 cases and 22 controls for study II). Informed consents of their parents or guardians were taken. They were subjected to clinical examination, relevant laboratory investigations, tuberculin test and chest radiograph. Relevant body fluids were subjected to bacteriological tests and PCR. Sera samples were analyzed for antibodies against PGLTbl and ESAT 6 antigen in study I and study II, respectively. ELISA tests showed a significantly higher sensitivity (49% study I; 53%, study II) as compared with LJ medium culture method (15.4%, study I; 28.9% study II) and ZN staining (27.7%, study I; 20.5%, study II) in all patients (p < 0.05). The results were comparable with PCR (40%, study I; 42.2% study II). Specificity of ELISA test was 100% in all the patients except in those with pulmonary disease (92.8%, study I; 84.8%, study II). In view of the convenience, low cost and comparable sensitivity with PCR, these ELISA tests have a promising future in the diagnosis of childhood tuberculosis.
Clinical and Vaccine Immunology, 2002
A radioassay for the detection of antitubercular antibody has been developed. The technique involves the addition of 125 I-labeled Mycobacterium tuberculosis antigen as a tracer, diluted clinical sample (serum or cerebrospinal fluid [CSF]), and heat-inactivated Staphylococcus aureus to capture the antibody, incubation for 4 h, and quantitation of the amount of antibody present in the sample. A total of 330 serum samples from patients with pulmonary tuberculosis and 138 control serum samples from individuals who were vaccinated with M. bovis BCG and from patients with pulmonary disorders of nontubercular origin were analyzed. Also, 26 CSF samples from patients with tuberculous meningitis and 24 CSF samples as controls from patients with central nervous system disorders of nontuberculous origin were analyzed. Sensitivities of 80 and 73% were observed for patients with pulmonary tuberculosis and tuberculous meningitis, respectively, and specificities of 90 and 88% were seen for the two groups of patients, respectively. The sensitivity was lower, however, for human immunodeficiency virus-infected patients coinfected with M. tuberculosis. The control population could be differentiated from the patient population. This assay is rapid and user friendly and, with its good sensitivity and specificity, should benefit the population by providing diagnoses early in the course of disease and, hence, permit the early administration of appropriate chemotherapy.
Serological diagnosis of tuberculosis
The Indian Journal of Pediatrics, 2008
To evaluate the efficacy of ELISA for the detection of IgG antibodies against antigen 85 complex (Ag 85 complex) of Mycobacterium tuberculosis.
Indian Journal of Clinical Biochemistry, 2001
Analysis of tubercular antibody, circulating free and immune complexed antigen (CIC-Ag) was done in confimed pulmonary tuberculosis sera by ELISA, using ES-31 antigen and affinity purified anti ES-31 antibody. Twenty three of 25 (92%) tuberculosis sera were positive for IgG antibody to ES-31 antigen. Using anti ES-31 antibody, free tubercular antigen could be detected in 20 of 25 (80%) cases whereas circulating immune complexed antigen (CIC-Ag) in 18 of 25 (72%) cases by sandwich ELISA. Of the two sera showing absence of antibody, one showed presence of free and CIC-Ag whereas the other showed the presence of free antigen. Thus antigen assay may be used as an adjunct tool for confirmation of pulmonary tuberculosis.
Diagnosis of Tuberculosis in Children: Increased Need for Better Methods
Emerging Infectious Diseases, 1995
In the last decade tuberculosis (TB) has reemerged as a major worldwide public health hazard with increasing incidence among adults and children. Although cases among children represent a small percentage of all TB cases, infected children are a reservoir from which many adult cases will arise. TB diagnosis in children usually follows discovery of a case in an adult, and relies on tuberculin skin testing, chest radiograph, and clinical signs and symptoms. However, clinical symptoms are nonspecific, skin testing and chest radiographs can be difficult to interpret, and routine laboratory tests are not helpful. Although more rapid and sensitive laboratory testing, which takes into account recent advances in molecular biology, immunology, and chromatography, is being developed, the results for children have been disappointing. Better techniques would especially benefit children and infants in whom early diagnosis is imperative for preventing progressive TB.