Evaluation of five rapid diagnostic tests for detection of antibodies to hepatitis C virus (HCV): A step towards scale-up of HCV screening efforts in India.

OBJECTIVES: Hepatitis C virus (HCV) infection is a major contributor to morbidity and mortality worldwide. Early detection and curative treatment of HCV can reduce the risk of liver-related mortality and serve to prevent transmission of new infections. India is estimated to have about six million HCV infected individuals, most of whom are unaware of their infection status. Rapid diagnostic test kits (RDTs) could help identify HCV infected persons more expeditiously and thus availability of high performing, quality-assured RDTs is essential to scale-up HCV screening efforts. The present study was thus undertaken to evaluate the performance characteristics of five anti-HCV RDTs. METHODS: Five anti-HCV RDTs (Alere Truline, Flaviscreen, Advanced Quality, SD Bioline and OraQuick) were evaluated using two panels of known anti-HCV positive and negative samples; one characterized from Indian patient samples (n = 360) and other obtained from the US Centers for Disease Control and Prevention (CDC), Atlanta (n = 100). Sensitivity, specificity, inter-observer agreement, test validity and operational characteristics of RDTs were assessed. RESULTS: The combined sensitivities across both panels for Alere Truline, Flaviscreen, Advanced Quality, SD Bioline and OraQuick RDTs were 99.4% (95%CI-96.6%-99.9%), 86.2% (95%CI-79.8%-91.1%), 96.2% (95%CI-91.9%-98.6%), 99.4% (95%CI-96.6%-99.9%) and 99.4% (95%CI-96.6%-99.9%) respectively. The overall specificities across both panels for all RDTs were 99.7%. The inter-observer agreement was 100% for Alere Truline, SD Bioline and OraQuick, while it was 99.5% and 98.6% with Advanced Quality and Flavicheck respectively. Discordant results were significantly associated with human immunodeficiency virus (HIV) positivity for both Advanced Quality and Flavicheck (p<0.001). CONCLUSION: The present evaluation demonstrated that Alere Truline, SD Bioline and OraQuick RDTs had sensitivity and specificity in accordance with the acceptance criteria of the Drug Controller General, India, the national regulatory authority, had excellent inter-observer agreement and superior operational characteristics. Our findings suggest that certain HCV RDTs perform well and can be a useful tool in screening of HCV infections expeditiously.

The Potential Impact of Up-Front Drug Sensitivity Testing on India's Epidemic of Multi-Drug Resistant Tuberculosis.

BACKGROUND: In India as elsewhere, multi-drug resistance (MDR) poses a serious challenge in the control of tuberculosis (TB). The End TB strategy, recently approved by the world health assembly, aims to reduce TB deaths by 95% and new cases by 90% between 2015 and 2035. A key pillar of this approach is early diagnosis of tuberculosis, including use of higher-sensitivity diagnostic testing and universal rapid drug susceptibility testing (DST). Despite limitations of current laboratory assays, universal access to rapid DST could become more feasible with the advent of new and emerging technologies. Here we use a mathematical model of TB transmission, calibrated to the TB epidemic in India, to explore the potential impact of a major national scale-up of rapid DST. To inform key parameters in a clinical setting, we take GeneXpert as an example of a technology that could enable such scale-up. We draw from a recent multi-centric demonstration study conducted in India that involved upfront Xpert MTB/RIF testing of all TB suspects. RESULTS: We find that widespread, public-sector deployment of high-sensitivity diagnostic testing and universal DST appropriately linked with treatment could substantially impact MDR-TB in India. Achieving 75% access over 3 years amongst all cases being diagnosed for TB in the public sector alone could avert over 180,000 cases of MDR-TB (95% CI 44187 - 317077 cases) between 2015 and 2025. Sufficiently wide deployment of Xpert could, moreover, turn an increasing MDR epidemic into a diminishing one. Synergistic effects were observed with assumptions of simultaneously improving MDR-TB treatment outcomes. Our results illustrate the potential impact of new and emerging technologies that enable widespread, timely DST, and the important effect that universal rapid DST in the public sector can have on the MDR-TB epidemic in India.

Catching the missing million: experiences in enhancing TB & DR-TB detection by providing upfront Xpert MTB/RIF testing for people living with HIV in India.

BACKGROUND: A critical challenge in providing TB care to People Living with HIV (PLHIV) is establishing an accurate bacteriological diagnosis. Xpert MTB/RIF, a highly sensitive and specific rapid tool, offers a promising solution in addressing these challenges. This study presents results from PLHIV taking part in a large demonstration study across India wherein upfront Xpert MTB/RIF testing was offered to all presumptive PTB cases in public health facilities. METHOD: The study covered a population of 8.8 million across 18 sub-district level tuberculosis units (TU), with one Xpert MTB/RIF platform established at each TU. All HIV-infected patients suspected of TB (both TB and Drug Resistant TB (DR-TB)) accessing public health facilities in study area were prospectively enrolled and provided upfront Xpert MTB/RIF testing. RESULT: 2,787 HIV-infected presumptive pulmonary TB cases were enrolled and 867 (31.1%, 95% Confidence Interval (CI) 29.4‒32.8) HIV-infected TB cases were diagnosed under the study. Overall 27.6% (CI 25.9-29.3) of HIV-infected presumptive PTB cases were positive by Xpert MTB/RIF, compared with 12.9% (CI 11.6-14.1) who had positive sputum smears. Upfront Xpert MTB/RIF testing of presumptive PTB and DR-TB cases resulted in diagnosis of 73 (9.5%, CI 7.6‒11.8) and 16 (11.2%, CI 6.7‒17.1) rifampicin resistance cases, respectively. Positive predictive value (PPV) for rifampicin resistance detection was high 97.7% (CI 89.3‒99.8), with no significant difference with or without prior history of TB treatment. CONCLUSION: The study results strongly demonstrate limitations of using smear microscopy for TB diagnosis in PLHIV, leading to low TB and DR-TB detection which can potentially lead to either delayed or sub-optimal TB treatment. Our findings demonstrate the usefulness and feasibility of addressing this diagnostic gap with upfront of Xpert MTB/RIF testing, leading to overall strengthening of care and support package for PLHIV.

Evaluation of Antioxidant and Hepatoprotective Activities of Moringa oleifera Lam. Leaves in Carbon Tetrachloride-Intoxicated Rats.

The antioxidant and hepatoprotective activities of the extract of Moringa oleifera leaves were investigated against CCl4-induced hepatotoxicity in rats. Hepatotoxic rats were treated with ethanol extract of Moringa oleifera for a period of 60 days at the following three dose levels; 100, 200 and 400 mg/kg body weight/day, orally. The activities were studied by assaying the serum marker enzymes like SGOT, SGPT, GGT, LDH, ALP, ACP, as well as total bilirubin, total protein and albumin in serum concomitantly with the activities of LPO, SOD, CAT, GSH, GR and GPx in liver. The activities of all parameters registered a significant (p ≤ 0.001) alteration in CCl4 treated rats, which were significantly recovered towards an almost normal level in rats co-administered with M. oleifera extract in a dose-dependent manner. All the biochemical investigations were confirmed by the histopathological observations and compared with the standard drug. silymarin. Results suggest that the antioxidant and hepatoprotective activities of M. oleifera leaves are possibly related to the free radical scavenging activity which might be due to the presence of total phenolics and flavonoids in the extract and/or the purified compounds ß-sitosterol, quercetin and kaempferol, which were isolated from the ethanol extract of M. oleifera leaves.

Antioxidant Potential of Plumieride against CCl4-Induced Peroxidative Damage in Rats.

In search of a new potent as an antioxidant from natural sources, plumieride-an iridoid isolated from the methanol extract of the bark of Plumeria bicolor (family Apocynaceae) was evaluated for its antioxidant potential against CCl4-induced peroxidative damage in liver of rats. The antioxidant potential was evaluated by using hepatic tissue for SOD (superoxide dismutase), CAT (catalase), GSH (reduced glutathione), GPx (glutathione peroxidase), GR (glutathione reductase) and LPO (lipid peroxidation) alongwith the concomitant blood serum for AST & ALT (aspartate and alanine transaminases), GGT (gamma glutamyl transpeptidase), ALP (alkaline phosphatase), total bilirubin and total protein contents. All the biochemical parameters were significantly (p ≤ 0.001) altered by CCl4 (0.3 mL/kg body weight/twice a week, intra-peritoneally for 30 days). Simultaneously, oral treatment with plumieride (5, 10 and 20 mg/kg body weight/day for 30 days), restored all the parameters towards a normal level, remarkably. The histological findings of liver sections further corroborated the antioxidant potential of plumieride compared with standard drug-silymarin. In conclusion, plumieride consists of sugar molecules, which have alcoholic groups. Therefore, the alcoholic groups of sugar increase its antioxidant potential through intermolecular hydrogen bonding along with the thiol(SH) group of non-protein thiols and enzymes resulting in the restoration of the antioxidant system. Therefore, it might be considered a natural antioxidant against peroxidative damage in rats.