B C Harinath's research while affiliated with Mahatma Gandhi Institute of Medical Sciences and other places
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Publications (189)
Background: Cholesterol degradation pathway is one of the important pathways in survival of Mycobacterium tuberculosis (Mtb) bacilli, and steroid C26-monooxygenase (CYP125) enzyme of Mtb associated with this pathway is reported to be novel drug target. This study aims to find out novel, safe, and effective inhibitors against CYP125 from natural phy...
Objective/background:
Tuberculosis (TB) has been declared as a global emergency by the World Health Organization in 1993 and still remains one of the world's biggest threats. Worldwide, 9.6 million people have been estimated to have fallen ill with TB in 2014: 5.4 million men, 3.2 million women, and 1.0 million children. To reduce this burden, det...
Isoniazid (INH), is one of the drugs shown to be effective and has been extensively used in TB control. Interestingly
tuberculosis showed predominant drug resistance to isoniazid and thus lead to multi drug therapy in TB treatment.
However, isoniazid is still advocated in latent TB and use as prophylactic in HIV infection and in children for preven...
![Schematic showing the activation and inactivation of INH [12]. INH is a...](profile/Lingaraja-Jena/publication/282533796/figure/fig2/AS:281004020387876@1444007987524/Schematic-showing-the-activation-and-inactivation-of-INH-12-INH-is-a-prodrug-and_Q320.jpg)
Objective/Background: Isoniazid (INH) is one of the effective antituberculosis (TB) drugs used for TB treatment. However, most of the drug-resistant Mycobacterium tuberculosis (MTB) clinical strains are resistant to INH, a first-line antituberculous drug. Certain metabolic enzymes such as adenosylhomocysteinase (Rv3248c), universal stress protein (...
Tuberculosis (TB) is an ancient disease caused by Mycobacterium tuberculosis (MTB), which remains a major cause for morbidity and mortality in several developing countries. Most drug-resistant MTB clinical strains are resistant to isoniazid (INH), a first-line anti-TB drug. Mutation in KatG, a catalase-peroxidase, of MTB is reported to be a major c...
Background:
Studies have been reported on genomic analysis to explain virulence of MTB H37Rv vs. MTB H37Ra strain. Proteomic comparison analysis at our centre has shown variability of 244 proteins in MTB H37Rv. A single amino acid mutation in protein sequence may cause alteration in protein structure and function that may account for virulence and...
Evaluation of SEVA TB ELISA using cocktail of mycobacterial antigens ES-31 and EST-6 (containing ES-38 and ES-41) and their specific antibodies in diagnosis of suspected cases of extrapulmonary tuberculosis. Detection of circulating free and immune complexed antigen to specific antibodies by sandwich ELISA and antibody to cocktail of antigens ES-31...
Background
Mycobacterium tuberculosis (MTB) H37Ra is an attenuated tubercle bacillus closely related to the virulent type strain MTB H37Rv. In spite of extensive study, variation in virulence between the MTB H37Rv and MTB H37Ra strains is still to be understood. The difference in protein expression or structure due to mutation may probably be an im...
Mycobacterial ES-31 serine protease has been reported to be a drug target using protease and lipase inhibitors in axenic and macrophage cultures. Simple screening techniques are needed for rapid testing of anti-tubercular drugs.
To demonstrate the usefulness of ELISA protocol based on antigenic reactivity of mycobacterial serine protease by indirec...
Objective
To assess the usefulness of in-house developed multiantigen and antibody assays, in diagnosis of both pulmonary and extra-pulmonary tuberculosis.Method
Clinically suspected cases of 31 pulmonary and 171 extra-pulmonary tuberculosis (TB) were screened by ELISA using cocktail (ES-31 + EST-6) antigen and their specific antibodies (anti ES-31...
Unlabelled:
MycoProtease-DB is an online MS SQL and CGI-PERL driven relational database that domiciles protease information of Mycobacterium tuberculosis (MTB) complex and Nontuberculous Mycobacteria (NTM), whose complete genome sequence is available. Our effort is to provide comprehensive information on proteases of 5 strains of Mycobacterium tub...
Isoniazid and orlistat were reported to have inhibitory effect on mycobacterial ES-31 serine protease in vitro and bacterial cell growth in axenic culture.
To study the cumulative effect and understand drug - drug interaction, if any, when isoniazid and orlistat used in combination.
Inhibition of mycobacterial ES-31 serine protease by different com...
Prospective evaluation of inhouse developed SEVA TB ELISA using cocktail of Mycobacterial antigens ES-31 and EST-6 (containing ES-38 and ES-41) and their specific antibodies in the diagnosis of Tuberculous pleural effusion was done in a tertiary care hospital.
Detection of circulating free and immune-complexed (IC) antigens and antibody by sandwich...
Tuberculosis (TB) is the leading cause of death worldwide attributable to a single infectious disease agent. India has more new TB cases annually than any other country. In 2008, India accounted for a fifth of the estimated 9.4 million TB cases globally. There is an overwhelming need for improving TB diagnostics in India through the use of cost eff...
The Mycobacterium tuberculosis Proteome Comparison Database (MTB-PCDB) is an online database providing integrated access to proteome sequence
comparison data for five strains of Mycobacterium tuberculosis (H37Rv, H37Ra, CDC 1551, F11 and KZN 1435) sequenced completely so far. MTB-PCDB
currently hosts 40252 protein sequence comparison data obtained...
Confirmation of presence of M. tuberculosis bacilli on microscopic examination is very important in diagnosis of tuberculosis. The present study was undertaken to find the usefulness of mycobacterial ES-31 serine protease as a marker to detect tuberculosis bacilli using fluorescein isothiocyanate conjugated anti-ES-31 serine protease antibody. This...
Mycobacterial excretory secretory-31 (SEVA TB ES-31) antigen is shown to possess protease and lipase activities.
To study the effect of commonly used HIV-protease inhibitors and lipase inhibitor Orlistat if any on mycobacterial ES-31 serine protease in vitro enzyme activity and on the growth of M.tb H37Ra bacilli in axenic culture.
Effect of HIV-pr...
Objective
To evaluate inhouse developed SEVA TB peroxidase enzyme immunoassay using cocktail of mycobacterial excretory-secretory antigens (ES-31, ES-43 & EST-6) for antibody detection and their affinity purified antibodies for antigen detection in tuberculosis suspected patients.Methods
Inhouse developed SEVA TB peroxidase enzyme immunoassay was e...
Decreased sensitivity has been a limiting factor of antigen assay for detection of tuberculosis. Assay of more than one antigen may improve sensitivity of an assay.
To develop a simple, rapid and less-expensive serodiagnostic method compared to culture method for Pulmonary Tuberculosis.
A cocktail of affinity purified antibodies against Mycobacteri...
There is a need for a simple and reliable method to identify Mycobacterium tuberculosis from nontuberculous mycobacteria (NTM).
The utility of mycobacterial ES-31, ES-43, EST-6 or ES-20 antigen as a biomarker for differentiation of Mycobacterium tuberculosis
bacilli from nontuberculous mycobacteria was explored using Fluorescein isothiocyanate conj...
There is a need for simple and reliable method to identify Mycobacterium tuberculosis from AFB smear positive cases. Utility of mycobacterial ES-31 serine protease as a marker to detect Mycobacterium tuberculosis bacilli was explored using Fluorescein isothiocyanate conjugated anti-ES-31 serine protease antibody. The presence of ES-31 serine protea...
To study the possible importance of mycobacterial ES-31 serine protease for bacterial cell growth, the effect of serine and metalloprotease inhibitors, anti-tubercular drugs such as isoniazid and anti-ES-31 antibody, was evaluated on mycobacterial ES-31 serine protease in vitro and on bacilli in axenic and macrophage cultures. Serine protease inhib...
SEVA TB Excretory secretory-31 (ES-31) antigen, a glycoprotein isolated from M. tb H37Ra culture filtrate, was found to be useful in the serodiagnosis of pulmonary tuberculosis (TB), extrapulmonary TB and in HIV-TB coinfection. Further, it has been shown to be a zinc containing serine protease.
To isolate and purify SEVA TB ES-31 antigen from M. tb...
An inhouse developed SEVA TB ELISA using cocktail of Mycobacterium Excretory-Secretory antigens (ES-31, ES-43 & EST-6) for antibody detection and their affinity purified antibodies for antigen detection was prospectively evaluated during January-August 2008 in TB suspected patients in tertiary hospital located in rural area. Antibody or antigen pos...
Immunoscreening was carded out by ELISA for the samples from the patients admitted in Kastruba Hospital, Sevagram. Blood samples of patients of pulmonary and extrapulmonary tuberculosis were examined for presence of tuberculosis specific antibody, antigen and combine sensitivity was also tested. Antigen preparation namely M.tbH 37R a ES-31 and 41 w...
Identification of in vitro and in vivo released mycobacterial antigens are of considerable interest in diagnosis of Mycobacterium tuberculosis. Isolation of in vitro released antigen from M. tb excretory-secretory culture filtrate protein and in vivo released circulating tuberculous antigen from smear positive pulmonary tuberculosis sera by ammoniu...
To determine role of antigens released in vivo and in vitro in immunodiagnosis of tuberculosis (TB).
In vivo released circulating tuberculosis antigen (CTA) was obtained from TB sera by ammonium sulphate precipitation and in vitro released excretory-secretory (ES) antigens from Mycobacterium tuberculosis culture filtrate. CTA and ES antigens were f...
Antigen assay is a better indicator of active infection than antibody detection hence affinity purified anti ES-31 antibody against Mycobacterium tuberculosis H 37 Ra Excretory-Secretory 31kDa serine protease antigen (SEVA TB ES-31) was explored for detection of circulating free and Immune-Complexed (IC) ES-31 antigen by microtitre plate Peroxidase...
Purpose: To determine role of antigens released in vivo and in vitro in immunodiagnosis of tuberculosis (TB). Methods: In vivo released circulating tuberculosis antigen (CTA) was obtained from TB sera by ammonium sulphate precipitation and in vitro released excretory-secretory (ES) antigens from Mycobacterium tuberculosis culture filtrate. CTA and...
Increased incidence of extrapulmonary tuberculosis (EPTB) as co-infection is observed due to rise in human immunodeficiency virus (HIV) infection. However, the precise diagnosis of EPTB is faced with difficulty due to the lack of tissue biopsy or fine-needle aspiration cytol- ogy (FNAC) facilities in rural hospitals. Enzyme linked immunosorbent ass...
Serological techniques like enzyme linked immunosorbent assay (ELISA) and immunoblotting are useful for detection of mycobacterial antigens of diagnostic importance in tuberculosis.
To isolate and identify circulating tuberculous antigens reactive with sputum positive and sputum negative pulmonary tuberculosis (PTB) sera.
Circulating tuberculous an...
Immunodiagnostically useful M. tuberculosis H37Ra protein antigens ES-31, ES-43 and EST-6 were isolated from detergent soluble sonicate (DSS) antigen using monospecific antibodies by affinity chromatography and compared with similar antigens isolated from M. tuberculosis culture filtrate for seroreactivity in tuberculosis sera by Indirect Enzyme Li...
To study tuberculous excretory-secretory (ES) 31 and ES-20 antigens in different pathogenic grades of lymph node tuberculosis (TB).
The study group included lymph node TB patients showing granuloma with mature epithelioid cells based on cytology findings (strong immune response group, SI) and patients showing no granuloma formation and acellular ne...
Lymphatic filarial subjects with disease manifestations exhibit significantly elevated levels of immune complexes (ICs) in their circulation. The objective of the study was to explore the possible excretion of filaria-specific soluble ICs in urine of subjects with lymphatic filariasis. Paired urine (overnight) and serum samples were analyzed for co...
In the present study we examined the utility of measuring antigen-specific soluble immune complexes (ICs) for the differential diagnosis and therapeutic monitoring of Bancroftian filariasis. Antigen-specific ICs were detected in significantly elevated levels in 90% of lymphatic filarial subjects suffering from disease manifestations, and very low l...
Mycobacterium tuberculosis excretory secretory 31 kDa, a serine protease antigen (M. tb ES-31), prepared from Mycobacterium tuberculosis H37Ra culture medium has been shown to have potential in detecting tuberculosis. Precise diagnosis and management of tuberculous meningitis, in children in particular, is essential to curtail mortality and morbidi...
A cocktail of affinity-purified antibodies against diagnostically useful Mycobacterium tuberculosis H37Ra excretory-secretory protein antigens ES-31, ES-43, and EST-6 was explored for detection of circulating free and immune-complexed (IC) antigen in sera of patients with confirmed tuberculosis (TB) by sandwich enzyme-linked immunosorbent assay and...
Lymphatic filariasis caused mainly by infection fromW. bancrofti andB. malayi remains a major cause of clinical morbidity in tropical and subtropical countries. Analysis ofB. malayi mf, infective larval and adult worm lysates for the activity of enzymes led to the demonstration of activities of three key enzymes of carbohydrate metabolism viz., Mal...
Monospecific antibodies have been successfully utilized in antigen detection, which is better indicator of active infection. Mycobacterium tuberculosis excretory secretory (M tb ES) antigens such as ES 31, ES 41 and ES 43 (31 kDa, 41 kDa and 43 kDa protein, respectively) have been shown to be present in Mycobacterium tuberculosis H37Ra culture filt...
To understand the usefulness of detecting tuberculous IgG antibodies against mycobacterial excretory-secretory 31 kDa serine protease antigen (SEVA TB ES-31) and circulating free and circulating immune-complexed (CIC) serine protease in TB patients with and without HIV infection.
Serum was collected from 144 individuals: patients with TB, with TB-H...
Clinical profile and serum beta-carotene levels in 100 cases of oral submucous fibrosis (OSMF) were studied. Prevalence of OSMF was noted to be 0.93% of new cases attending ENT OPD Age of cases varied from 12 to 78 years (mean 29.09 years) with male-female ratio of 3.3:1 and 80% literacy. Students constituted the single largest group. Burning sensa...
The seroreactivity of a cocktail of purified mycobacterial excretory-secretory (ES) antigens ES-31, ES-41 and ES-43 was assessed by ELISA and immunoblotting in patients with pulmonary tuberculosis. The ES-31 antigen was isolated by affinity chromatography and ES-41 and ES-43 were isolated by fast protein liquid chromatography from Mycobacterium tub...
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...
Glutathione-S-transferase (GST) has been detected in the adult female Setaria cervi, a bovine filarial parasite. The role of S. cervi GST antigen in inducing immunity in the host against Brugia malayi microfilariae and infective larvae was studied by in vitro antibody dependent cell mediated reaction as well as in situ inoculation of filarial paras...
A 175 kDa antigen fraction with collagenase activity was isolated and purified from somatic extracts of adult Setaria cervi females using column chromatography involving consecutive steps of DEAE-Sepharose CL6B and Sephadex G-100. The optimum pH for 175 kDa collagenase was found to be pH 7.0. Sensitivities to a variety of inhibitors and activators...
An Excretory Secretory protein antigen of 6 kDa (ES-6) was isolated from Mycobacterium tuberculosis H 37Ra culture filtrate by gel filtration using fast protein liquid chromatography. Seroreactivity of ES-6 antigen was compared with earlier reported diagnostically useful ES-31 and ES-43 antigens at different stage of pulmonary tuberculosis and in h...
Seroreactivity of bancroftian filarial cases of different endemic zones with different geogrophical isolates of W.bancrofti mf was checked by indirect fluorescent antibody test (IFAT). While the filarial sera of Calicut showed binding to the mf isolates of all four endemic zones studied (Calicut, Bhubaneshwar, Rourkela and Wardha), the sera of Bhub...
The ES-31 (31 kDa protein) antigen was isolated from culture filtrate of Mycobacterium tuberculosis H37Ra and was shown to have potential in immunodiagnosis of pulmonary tuberculosis. Serum samples from 38 confirmed sputum positive pulmonary tuberculosis patients were grouped into AFB+, AFB++, AFB+++ based on sputum bacillary load. Analysis of tube...
The secreted protein antigens provide first stimulus in vivo for humoral and cellular immune response to mycobacteria and may play useful role in serodiagnosis. A secretory protein ES-31 isolated from Mycobacterium tuberculosis H37Ra culture filtrate has been earlier shown diagnostically useful in pulmonary tuberculosis. Similarly ESAS-6 antigen fo...
Lipid peroxidation product, malonaldehyde (MDA) and antioxidants were estimated in plasma and erythrocytes of 34 cases of oral submucous fibrosis (OSMF) of different grades with equal number of healthy controls to evaluate the association of reactive oxygen species (ROS) and OSMF. While plasma MDA was found to be significantly higher in patients (3...
Serodiagnosis by ELISA has been widely explored over the years, in the diagnosis of tuberculosis. Two ELISA systems were evaluated for detection of mycobacterial antibodies in pulmonary and extra pulmonary tuberculosis. The two test assays explored were ERBA LISA (TB IgG) test (Anda Biologicals) which uses A60 antigen complex found in the cytosol o...
Tuberculosis is still a major health problem in most developing countries and its incidence is rising in many developed countries. This resurgence has been attributed to the HIV epidemic and TB has been declared as a global health emergency by WHO in 1993. The diagnosis of tuberculosis mainly depends upon initial clinical suspicion and radiographic...
Despite rapid advances in molecular genetics for detection of mycobacteria, it is clear that interest in serodiagnosis remains high, especially for those situations in which a specimen may not contain the infecting agent in particular in extrapulmonary tuberculosis. Immune response to excretory-secretory (ES) proteins of Mycobacterium tuberculosis...
To isolate and characterise in vivo released 41 kDa mycobacterial antigen in pulmonary and bone and joint tuberculosis (TB) and its identification with in vitro released ES-41 kDa antigen.
Circulating antigen was isolated from confirmed pulmonary tuberculosis serum (PTS) and bone and joint tuberculosis serum (BJS) by trichloroacetic acid precipitat...
Analysis of immune response in individuals with different clinical manifestations living in filaria endemic area will be of interest to understand the immunological events associated with the disease development in filaria infected endemic population. The levels of four IgG subclasses and IgE antibodies against Brugia malayi microfilarial excretory...
Sera samples of 7 microscopic haematuria cases collected before and after treatment with Diethylcarbamazine citrate, (DEC), 9 microfilaraemic cases and 19 endemic normal individuals were analysed for filarial antigen and IgG antibody levels. Filarial antigen was detected in 5 of the 7 microscopic haematuria cases, of which 3 turned negative for ant...
A serological test with good diagnostic potential in pulmonary and in particular extra pulmonary tuberculosis, a paucibacillary condition would contribute to early detection and help in better management of patients. Two different secretory proteins of Mycobacterium tuberculosis (M. tb) H37Ra viz; ES-31 with diagnostic potential in pulmonary TB, tu...
Tuberculosis remains major health problem in India and developing countries Immunodiagnosis has important role in screening, diagnosis and management of tuberculosis. SEVA TB ES-31 antigen has shown potential in detecting tuberculous IgG antibody in earlier studies from our laboratory. In the present study we have analysedSEVA TB ES-31 antigen spec...
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 antig...
A comparative analysis was made on the utility of SEVAFILACHEK-stick based immunoassays and commercially available ICT-filariasis test to detect active infection in different groups of bancroftian filariasis. The SEVAFILACHEK immunoassays were found to be useful to detect filarial infection in microfilaraemia and in a significant number of clinical...
Excretory-secretory proteins of Mycobacterium tuberculosis H37Ra, have been of diagnostic interest in pulmonary (PTB) and extrapulmonary tuberculosis (EPTB). Two different excretory-secretory antigenic proteins of M.tbH37Ra viz., EST-DE1 (a 6% TCA soluble and DEAE anion exchange purified antigen) and ESAS-7 (50% ammonium sulphate solubilized and SD...
Proteins secreted into the culture medium by Mycobacterium tuberculosis are shown to be a source of antigens of immunodiagnostic importance. In our earlier study, we had reported a 31-37 kDa seroreactive gel-eluted antigenic fraction (ESAS-7), isolated from culture filtrate proteins of Mycobacterium tuberculosis H37Ra. In this report, we describe f...
Proteins secreted into the culture medium byMycobacterium tuberculosis (M. tb) are shown to be source of antigens of immunodiagnostic interest. Anin vitro released 31 kDa antigen ESAS-7F isolated fromM.tb H37Ra culture filtrate by salt precipitation, SDS-PAGE and cation exchange fast protein liquid chromatography (FPLC) was shown
earlier to be a di...
Lymphatic filariasis is a major public health problem in India with 412 million people living in bancroftian endemic areas and is a major cause of clinical morbidity. Twenty million people are reported to suffer from chronic disease manifestations such as lymphoedema, hydrocele or elephantiasis. At least twice the number have been shown to suffer f...
Brugia malayi microfilarial excretory-secretory (mf ES) and phosphate buffer saline soluble (mf S) antigens were fractionated by fast protein liquid chromatography (FPLC) on superdex 200 HR 10/30 gel filtration column. The active antigen fractions were identified and explored in comparison with whole mf ES and mf S antigens to detect filarial IgG a...
Oxidative stress was studied by estimating plasma levels of malondialdehyde (MDA), beta carotene, vitamin E and erythrocytic superoxide dismutase(E-SOD) activity in 50 cases of carcinoma of upper digestive tract which included carcinoma of oral cavity, pharynx and oesophagus. While plasma MDA level was found to be increased (3.5±1.0 nmole/ml), a si...
Lymphatic filariasis caused mainly by infection fromWuchereria bancrofti andBrugia malayi remains as the major cause of clinical morbidity in tropical and subtropical countries. Development of vaccine against filarial infection can act as additional measure to the existing therapeutic and vector control methods in the control of this disease. The m...
Trichloroacetic acid (TCA) solubilized and DEAE fractionatedMycobacterium tuberculosis H(37)Ra excretory-secretory (ES) antigen viz., Mtb EST DE1 and affinity purified goat antibodies to the TCA solubilized ES antigen (Mtb EST) were explored in detecting tubercular antibody and antigen respectively in sera of bone and joint tuberculosis by indirect...
Antigens released in vivo are of considerable interest in the immunodiagnosis of infectious diseases. Circulating antigen was isolated from bacteriologically confirmed tuberculous sera by ammonium sulphate precipitation. The protein fraction between 36%, and 75%, ammonium sulphate was reactive with tuberculosis (TB) sera showing the presence of cir...
Filariasis is a major public health problem in India. About 412 million people are living in bancroftian endemic areas distributed in 13 states and 5 union territories. Rural areas have the population at risk three times more than the urban India. The surveys in 1994 have shown that 26.9 million people are harbouring Wuchereria bancrofti microfilar...
Brugia malayi microfilarial excretory-secretory (mf ES) antigens obtained byin vitro maintenance of mf are important tools in the immunodiagnosis of bancroftian filariasis. To increase the yield of mf ES products,
the effect of nutritional supplements on the culture medium (RPMI 1640) and the maintenance temperature were studied. Supplementation
of...
Lymphatic filariasis continues to be the major cause of clinical morbidity in India and other developing tropical countries.
One of the major lacunae in the effective management of clinical filarial cases is the non-availability of a suitable diagnostic
test for confirming filaria aetiology in acute, chronic and occult clinical cases where microfil...
A clinical study and immunoscreening was conducted on 363 suspected filarial patients attending the surgery out patient division of the MGIMS, Sevegram. The disease was significantly higher in males (86%) than in females (14%). Majority (52.9%) of the cases were in the age group of 11-30 years. The distribution of cases into three different grades...