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M. V. R. Reddy R. Alli B. C. Harinath 《Indian journal of clinical biochemistry : IJCB》2000,15(1):127-135
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 main hurdles in the development of anti-filarial vaccine are the strict primate specificity ofWuchereria bancrofti, the paucity of parasite material, the diversity of clinical manifestations and their associated complex immune responses, lack of clear understanding on host-parasite interactions and the mechanisms involved in protective immunity. However in the past few years, the information generated in immuno-epidemiological studies, correlated with observations in experimental animals suggests that a filarial vaccine is feasible. Initially live irradiated infective larvae have been successfully used to induce high level of protective immunity in several animal models. Applying diverse strategies, variety of purified or recombinant filarial antigens have been explored for their ability to induce protection in different host-parasite systems. Some of these targeted filarial antigens induced high level of resistance in experimental animals against challenge infections. More focussed studies on thorough characterization of parasitological and immunological changes associated with resistance induced by such candidate protective antigens and on delivery mechanisms and safety aspects will be crucial in their selection for possible use in humans. 相似文献
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Sonika Gupta Niraj Shende Swati Banerjee Satish Kumar M. V. R. Reddy Bhaskar C. Harinath 《Indian journal of clinical biochemistry : IJCB》2002,17(1):5-8
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 specific immunoglobulinsIgM, IgA and IgG in clinically and bacteriologically confirmed pulmonary tuberculosis cases to determine the usefulness of specific immunoglobulin
class in the diagnosis of patients attending the hospital.
Of the 30 cases of pulmonary tuberculosis 25 (83.3%) were positive for IgG, 19 (63.3%) for IgM and 16 (53.3%) for IgA. On
combining IgG and IgM positivity, sensitivity was increased to 93.3%. While combining IgG and IgA positivity, sensitivity
increased to 90%. However specificity was decreased to 66.6% and 70% for both of these combinations respectively. It could
be envisaged from this study that IgG antibody detection against ES-31 antigen showed acceptable sensitivity (83.3%) and specificity
(86.6%) compared to IgM or IgA alone or in combination. When immune responses were analysed according to degree of sputum
positivity, IgG response was observed to be predominant in all grades, compared to IgM or IgA antibody. The addition of IgM
or IgA as an adjunct test increases the sensitivity but at the cost of specificity. Hence the detection of IgG alone is more
useful compared to IgM or IgA assay, in detecting tuberculosis disease cases coming to the hospital. 相似文献
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Y. P. Bhandary K. N. Krithika Sandeep Kulkarni M. V. R. Reddy B. C. Harinath 《Indian journal of clinical biochemistry : IJCB》2006,21(1):1-7
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., Malate dehydrogenase (MDH), Malic enzyme (ME) and Glucose-6-phosphate dehydrogenase
(G6PDH) in all the three stages of the parasite. The specific activity of all the three dehydrogenases was significantly high
in mf lysate compared to their activity in lysates of the other two stages (P<0.001). Analysis by native polyacrylamide gel
to their activity inlysates of the other two stages (P<0.001). Analysis by native polyacrylamide gel electrophoresis (PAGE)
using 7.5% non-gradient gel showed the presence of two isoforms of each of the three enzymes (MDH, ME & G6PDH) in mf lysate,
while only one form of each enzyme was present in L3 larval and adult worm lysates. Further proteolytic enzyme activity was demonstrated both in microfilarial and infective larval
lysates ofB. malayi. While both mf and L3 larval lysates showed optimal protease activity at alkaline pH of 9.0, the mf lysate showed increased activity also at pH
3.0. The infective larval lysate was markedly inhibited by Tosylamide-L-Phenylalanine chloromethyl ketone (TPCK), a thiol
protease inhibitor, while the protease activity in mf lysate was significantly inhibited by both TPCK and a serine protease
inhibitor Phenyl Methyl Sulphonyl Flouride (PMSF). In sodium do-decyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE),
using gelatin copolymerized gel, the microfilarial lysate showed 3 protease molecules of 40 kDa, 180 kDa and 200 kDa and the
L3 larval lysate had 6 protease molecules of 18, 25, 37, 49, 70 and 200 kDa size. 相似文献
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R. Alli B. Bhunia G. P. Chhotray M. V. R. Reddy B. C. Harinath 《Indian journal of clinical biochemistry : IJCB》2003,18(1):61-64
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 antigen after treatment with
DEC. While none of the 7 haematuria cases were positive for filarial IgG antibodies, before the DEC treatment, all of them
turned positive after DEC treatment. The sensitivity and specificity values(to detect mf +ve cases) were 89% and 90% respectively
for the detection of filarial antigen and 78% and 95% respectively for the detection of filarial IgG antibodies. 相似文献
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J. Pramanik A. N. Lodam C. M. Badole M. V. R. Reddy K. R. Patond B. C. Harinath 《Indian journal of clinical biochemistry : IJCB》2000,15(1):22-28
Trichloroacetic acid (TCA) solubilized and DEAE fractionatedMycobacterium tuberculosis H37Ra 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 and sandwich ELISA. Out of total 36 bone & joint tuberculosis cases, tubercular antibody was detected by indirect
ELISA in 30 patients (sensitivity 83%), while circulating tubercular antigen was detected by sandwich ELISA in 27 patients
(sensitivity 75%). Out of 34 non tubercular disease control cases, 10 patients showed positive reaction for antibody while
only 4 patients showed positive reaction for antigen. In another group of 34 healthy subjects who were screened, 4 individuals
showed positive reaction for tubercular antibody and 2 cases for antigen. This study shows that antigen detection assay using
affinity purified anti Mtb EST antigen antibody is superior with overall specificity of 91% as compared to antibody detection
assay with 75% specificity in bone & joint tuberculosis. 相似文献
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Soma Gupta K. K. Singh V. J. Vyas V. N. Chaturvedi M. V. R. Reddy B. C. Harinath 《Indian journal of clinical biochemistry : IJCB》2000,15(1):52-55
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 significant decrease in beta
carotene (81.2±14.5mg%), vitamin E (8.5±1.1 mg/L) level and E-SOD activity (657.0±80.6 U/G Hb) were observed in carcinoma
of upper digestive tract. Patients were treated with radiotherapy which itself was toxic enough and produced its deleterious
effects by generation of reactive oxygen species (ROS). As antioxdiants can detoxify ROS, beneficial effect if any, of antioxidant
administration during radiotherapy was studied in two groups of patients, group A (n=5, supplemented with antioxidants) and
group B (n=5, without antioxidant supplementation). Plasma MDA level was found to be elevated in both the groups but the increase
in group B was significant, compared to pretreatment level. Further, body weight was found to be significantly decreased in
group B patients, which was maintained in group A patients. Moreover, group A patients showed significant elevation in beta
carotene concentration, thus showing beneficial effect of administration of antioxidants during radiotherapy without disturbing
the desirable therapeutic effect of radiotherapy. 相似文献
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B. C. Harinath M. V. R. Reddy B. Bhunia Y. P. Bhandari V. K. Mehta P. Chaturvedi N. C. Prajapati R. K. C. Gupta 《Indian journal of clinical biochemistry : IJCB》2000,15(1):118-126
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 from acute and occult filarial infections in an endemic area without diagnosis. Due to non-availability of suitable diagnostic test for confirming filaria aetiology other than parasitological examination, no significant study on filariasis in children has been reported earlier. Studies in our laboratory for more than a decade showed usefulness of microfilarial excretory-secretory antigen in confirming filarial aetiology in acute and occult infections in adults as well as in children. This study reports acute and atypical manifestations such as lymphadenopathy, asthmatic bronchitis, pulmonary eosinophilia, mono-arthritis, recurrent URI, pneumonia, nutritional anemia, pain in abdomen etc. in children living in filaria endemic area having no microfilaraemia but showing filaria aetiology by immunomonitoring for the presence of antibody or antigen and responding to optimal DEC therapy. 相似文献
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E. Raji Nair Swati Banerjee Satish Kumar M. V. R. Reddy B. C. Harinath 《Indian journal of clinical biochemistry : IJCB》2001,16(1):132-135
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 diagnostically important antigen fraction. In this report, we describe the isolation of ESAS-7F antigen using
monospecific antibody coupled to sepharose CL-4B column. The percentage recovery of ESAS-7F antigen using affinity chromatography
was approximately 8% of the total ES antigen proteins compared to 0.05% obtained by conventional purification steps using
salt precipitation, SDS-PAGE and FPLC. Similar seroreactivity was observed by the antigen isolated by both the methods in
indirect ELISA. Affinity chromatography helped in an increased recovery of ESAS-7F antigen and obviates the need for time
consuming conventional purification steps. 相似文献