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Biomedical subjects

V Lakshmi

Publications and source records attributed to V Lakshmi.

At least 19 recordsLinked to original sources

Rapid and real-time detection of Chikungunya virus by reverse transcription loop-mediated isothermal amplification assay.

The standardization and validation of a one-step, single-tube, accelerated, quantitative reverse transcription (RT) loop-mediated isothermal amplification (RT-LAMP) assay targeting the E1 gene for the rapid and real-time detection of Chikungunya virus (CHIKV) are reported. A linear relationship between the amount of template and time of positivity value over a range of 2 x 10(8) to 2 x 10(2) copies was obtained. The feasibility of CHIKV RT-LAMP for clinical diagnosis was validated with patient serum samples from an ongoing epidemic in Southern India. Optimal assay conditions with zero background were established for the detection of low levels of CHIKV in acute-phase patient serum samples. The comparative evaluation of the RT-LAMP assay with acute-phase patient serum samples demonstrated exceptionally higher sensitivity by correctly identifying 21 additional positive borderline cases that were missed by conventional RT-PCR (P < 0.0001) with a detection limit of 20 copies. The quantification of virus load in patient serum samples was also determined from the standard curve based on their time of positivity and was found to be in the range of 2 x 10(8) to 2 x 10(1) copies. In addition, the field applicability of the RT-LAMP assay was also demonstrated by standardizing SYBR Green I-based RT-LAMP wherein the amplification was carried out in a water bath at 63 degrees C for 60 min, which was followed by monitoring gene amplification with the naked eye through color changes. These findings demonstrated that the RT-LAMP assay is a valuable tool for rapid, real-time detection as well as quantification of CHIKV in acute-phase serum samples without requiring any sophisticated equipment and has potential usefulness for clinical diagnosis and surveillance of CHIKV in developing countries.

Acute Disease↗

Antifertility activity of Piper longum Linn. in female rats.

The crude extract, its different fractions and the major pure compound from the active fraction of the powdered fruits of Piper longum were studied for the antifertility effect in female rats. The crude extract and its hexane fraction exhibited 100 and 86% efficacy respectively (days 1-7 post-coitum (p.c.) schedule). On the other hand, 1-butanol soluble, 1-butanol insoluble and chloroform fractions were inactive.

Abortifacient Agents↗

Identification and characterization of a major cell wall-associated iron-regulated envelope protein (Irep-28) in Mycobacterium tuberculosis.

Iron limitation and the expression of mycobactin and carboxymycobactin by Mycobacterium tuberculosis are known. Here, we report how iron regulated the coordinate expression of these two siderophores and a 28-kDa cell wall-associated iron-regulated protein (Irep-28). Irep-28 is identified as the DNA-binding HU homologue HupB protein (hupB [Rv2986c]). Antibodies to this protein were detected in sera from tuberculosis patients. The location of the protein in the cell wall makes it a potential drug target.

Amino Acid Sequence↗

Changing trends in the antibiograms of Salmonella isolates at a tertiary care hospital in Hyderabad.

The present study was undertaken to compare the changing trends of antibiograms of Salmonella enterica serovar Typhi and Salmonella enterica serovar Paratyphi A isolates. A total of 80 isolates of salmonella obtained from blood cultures between 2001-2004 were included in the study. Identification and antibiotic sensitivities of the isolates were performed by using mini API (bio Merieux, France). Sixty isolates were identified as Salmonella enterica serovar Typhi and 20 were identified as Salmonella enterica serovar Paratyphi A. More than 67% of S.typhi and 80% of S.paratyphi A isolates were sensitive to chloramphenicol. Sensitivity of S.typhi isolates to cephalosporins was found to have increased from 2001-2004 while that of S.paratyphi A showed a decline. With increasing resistance to ciprofloxacin and the possibility of re-emergence of sensitivity to chloramphenicol, the policy of empirical treatment of enteric fever needs to be rationalized.

Anti-Bacterial Agents↗

Ventriculoperitoneal shunt infections.

Central nervous system (CNS) shunt infection is a cause of significant morbidity, causing shunt malfunction and chronic ill health. This study was carried out to evaluate the infection rate associated with CNS shunts, assess the frequency of the pathogens as well as their antibiotic sensitivity pattern aiming at suitable prophylaxis. A retrospective analysis of 226 CSF cerebrospinal fluid (CSF) shunt procedures sent for bacteriological work up over a period of one year and six months was undertaken. Laboratory diagnosis was established by subjecting the CSF to cell count, biochemical tests, bacteriological culture and antibiotic susceptibility test. Nine out of 226(3.98%) of the CSF samples were culture positive. Coagulase negative Staphylococcus was the most common isolate accounting for 36.36%. Majority of the isolates were sensitive to the thirdgeneration cephalosporins and quinolones. The antibiotic sensitivity pattern suggests cephalosporins and quinolones to be a better choice of antibiotics either prophylactically or therapeutically, which may result in effective and rapid sterilisation of the CSF.

Adolescent↗

Utility of HCV core antigen ELISA in the screening for hepatitis C virus infection in patients on hemodialysis.

An enzyme immuno assay for hepatitis C core antigen was recently developed and its performance was compared with that of the hepatitis C virus (HCV) RNA in the screening of HCV infection in patients on hemodialysis. One hundred and eleven chronic renal failure patients undergoing haemodialysis between May 2003 and October 2004 were included in the study. All the patients were tested for anti HCV antibody, core antigen and RNA. Fifteen patients were anti HCV antibody positive, three patients were positive for HCV core antigen and RNA, three patients were positive for HCV RNA, while two patients were positive only for core antigen but negative for RNA. In anti HCV antibody positive patients, the core antigen was negative while the viral RNA continued to be present. Hence, relying solely on a single HCV core antigen assay may not be useful for a definite diagnosis of early HCV infection. The sensitivity and specificity of the assay were 60% and 83% respectively, while the positive predictive value was 14.3%, negative predictive value was 97.7% and the efficiency was 81.9%.

Enzyme-Linked Immunosorbent Assay↗

Isolation of mycobacteria by BACTEC 460 TB system from clinical specimens.

This article reports our experience with the BACTEC 460 TB system in the past five years and its performance characteristics and its advantages over the conventional LJ medium for mycobacterial culture. Clinical specimens (3597) from patients suspected to have tuberculosis were submitted for mycobacterial culture between May 2000 and August 2005 and were processed using the BACTEC 460 TB system. Pulmonary samples were 1568 while the extra pulmonary samples were 2029. BACTEC achieved detection of 681 (18.93%) M. tuberculosis cases (499- pulmonary, 182- extrapulmonary) with a recovery time shorter by 13.2 days compared to conventional method, while 577 (84.7%) were non-tuberculosis mycobacteria. Automated systems can have a great impact and thrust on an early diagnosis of tuberculosis allowing an early and appropriate management of the patient and thereby a better disease outcome.

Automation↗

The role of quantitative cultures of non-bronchoscopic samples in ventilator associated pneumonia.

PURPOSE: The objective of this study is to determine the role of quantitative cultures of non-bronchoscopic samples such as blinded bronchial sampling (BBS) and endotracheal aspirates (ETA) in the management of ventilator associated pneumonia (VAP). The study also evaluates the clinical diagnosis of VAP based on the inclusion of Gram stain results of BBS/ETA samples into modified clinical pulmonary infection score (CPIS). METHODS: Fifteen out of the 120 patients admitted to respiratory intensive care unit under mechanical ventilation for more than 48 hours with a clinical suspicion of VAP, were included in this study. Quantitative cultures of BBS and ETA were performed from all the 15 patients. RESULTS: VAP was confirmed in 11 out of 15 cases by quantitative cultures of either the BBS or ETA samples. The condition of 8/11 VAP confirmed patients improved significantly with the change in antibiotic therapy. The overall mortality rate was found to be 18%. The agreement between BBS and ETA results was found to be 83.3%. Modified-clinical pulmonary infection score (CPIS) increased significantly when Gram stain results of BBS/ETA samples were included, thereby strengthening the clinical diagnosis of VAP. CONCLUSIONS: Quantitative culture of lower respiratory tract samples obtained by non-bronchoscopic methods may be a useful alternative to bronchoscopy, in the diagnosis of VAP. Inclusion of Gram stain results of BBS/ETA into modified-CPIS may augment the diagnostic evaluation of VAP.

Bacteria↗

Serodiagnosis of dengue virus infection in patients presenting to a tertiary care hospital.

Dengue is an acute infectious disease of viral etiology. It is probably one of the most important arthropod borne viral disease in terms of human morbidity and mortality. The spectrum of disease ranges from self-limited dengue fever to more severe forms of dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS). Laboratory diagnosis of dengue virus infection mainly depends on detection of virus specific antibodies. The aim of the study was to correlate the serological results with clinical presentation in patients with a diagnosis of dengue. Eleven out of 15 (73.3%) patients with DHF and DSS had secondary antibody response and mortality was 100% in these patients.

Academic Medical Centers↗

Occurrence of extended spectrum beta-lactamases among Enterobacteriaceae spp. isolated at a tertiary care institute.

Increasing resistance to third generation cephalosporins has become a cause for concern especially among Enterobacteriaceae that cause nosocomial infections. The prevalence of extended spectrum beta-lactamases (ESBLs) among members of Enterobacteriaceae constitutes a serious threat to current beta-lactam therapy leading to treatment failure and consequent escalation of costs. A detailed study was initiated to identify the occurrence of ESBLs among the Enterobacteriaceae isolates at a tertiary care hospital using the double disk potentiation technique. Antibiogram profiles were determined to commonly used antibiotics and confirmation of ESBLs production was carried out by the disk diffusion assay using ceftazidime and cefotaxime in the presence and absence of clavulanic acid. Our results indicate that the majority of ESBLs were expressed in Escherichia coli.

Anti-Bacterial Agents↗

Pulmonary zygomycosis in a diabetic patient.

We report a case of pulmonary zygomycosis in an adult male diabetic patient who presented with fever and altered sensorium initially and later developed streaky haemoptysis. Bronchoscopy showed picture of necrotizing pneumonia. Sputum was negative for fungal elements on admission but later bronchial wash and repeat sputum samples were positive by microscopy and culture showed growth of Rhizopus species. Immediately the patient was put on amphotericin B but had a bout of massive haemoptysis and succumbed. A high index of suspicion is needed for an early diagnosis and aggressive treatment of this infection in view of the high mortality rate.

Amphotericin B↗

Pathogenesis and pathology of brain abscess.

Brain abscess continues to be a serious medical problem with increasing incidence despite advances in diagnostic and surgical methods, and advent of new antibiotics. This is due to increase in immune suppressed individuals, opportunistic pathogens and resistance to antibiotics. The morbidity, mortality and long term sequelae of brain abscess like cognitive impairment and poor neurological outcome are due to persistent release of proinflammatory mediators by activated microglia, astrocytes and infiltrating inflammatory cells, along with disruption of blood brain barrier. Anti-inflammatory drugs along with specific antimicrobial agents help in minimizing damage to the adjacent brain parenchyma. Identification of microorganisms in the aspirated material or the excised specimen help decide the optimal antimicrobial therapy. Prompt examination of smear and meticulous culture techniques to identify aerobes, anerobes, mycobacteria, actinomycetes, parasites and fungi are of utmost importance in choosing antibiotics. Histology complements culture and plays a key role especially in specific infections. Use of appropriate histochemical stains along with tissue reaction helps in diagnosis. The route of spread, the type and virulence of the organism, thickness of the capsule, location and number of abscesses in the brain, and immune status of the host are important determinants of outcome. Identification of microorganisms and insights into pathogenesis allow appropriate therapeutic interventions to improve outcome.

Actinobacteria↗

In vitro and in vivo leishmanicidal activity of Dysoxylum binectariferum and its fractions against Leishmania donovani.

The leishmanicidal effect of crude ethanolic extract of stem bark of Dysoxylum binectariferum and its fractions has been investigated against Leishmania donovani, the causative agent of visceral leishmaniasis. Ethanolic extract was lethal to promastigotes as well as amastigote forms in macrophage system at the concentration of 100 microg/ml. Chloroform fraction significantly inhibited promastigote multiplication and was also active against amastigotes in infected J774A.1 macrophages at 100 microg/ml. Hexane fraction was moderately active and the other fractions were inactive against both the forms. When tested in vivo in hamsters, ethanolic extract was toxic at 500 mg/kg whereas exhibited marginal activity (67.7+/-5.3%) at 250 mg/kg x 5, p.o. on day 7 post treatment (p.t.) which increases slightly (69+/-4.7) by day 30 p.t. Chloroform and n-hexane fractions exhibited 64.3+/-4% and 47.8+/-4.6% parasite inhibition at the dose of 100 mg/kg x 5 p.o., respectively. The pure compound, rohitukine, obtained from chloroform fraction showed weaker in vitro activity and was ineffective in infected hamsters. The lead potential of this plant need further investigations.

Animals↗

Comparative evaluation of TRI-DOT Rapid HIV test with fourth-generation ELISA for the detection of human immunodeficiency virus.

This study evaluated the TRI-DOT Rapid HIV test for the early detection of human immunodeficiency virus (HIV) infection in comparison with a fourth-generation ELISA (Vironostika HIV Uniform II). Of 23,609 sera, seven (0.03%) gave discordant results. Six of these were reactive only by the fourth-generation assay and were p24 antigen-positive by VIDAS DUO, Western blot and qualitative RT-PCR tests. The remaining discordant serum was considered to be false-positive by the TRI-DOT assay, as it was negative by repeat ELISA and Western blot tests. The sensitivity and specificity of the TRI-DOT test were 99.48% and 99.99%, respectively, compared with the fourth-generation ELISA.

AIDS Serodiagnosis↗