Investigating the dynamics of Leishmania antigen in the urine of patients with visceral leishmaniasis: a pilot study

Background: Detection of Leishmania antigens in the urine provides a non-invasive means of diagnosis and treatment monitoring of cases of visceral leishmaniasis (VL). Leishmania antigen load in the urine may vary between different time-points within a day, thus influencing the performance of antigen-detection tests. Methods: We investigated the dynamics of Leishmania antigen in urine collected at three different time points (08:00, 12:00 and 16:00 hours). All urine samples collected were tested with the Leishmania Antigen ELISA (VL ELISA) kit, produced by Kalon Biological Ltd., UK. Results: The median concentration of Leishmania antigen in urine collected at 08:00 (2.7 UAU-urinary antigen units/ml) was higher than at 12:00 (1.7 UAU/ml) and at 16:00 (1.9 UAU/ml). These differences were found to be statistically significant (08:00 vs. 12:00, p=0.011; 08:00 vs. 16:00, p=0.041). Conclusion: This pilot study indicates that the Leishmania antigen concentration is higher in urine samples collected in the morning, which has important implications when the VL ELISA kit or other tests to detect Leishmania antigen in urine are used for diagnosis of VL and treatment monitoring.

Download Full-text

Testing urine samples with rK39 strip as the simplest non-invasive field diagnosis for visceral leishmaniasis: An early report from eastern India

Journal of Postgraduate Medicine ◽

10.4103/0022-3859.101378 ◽

2012 ◽

Vol 58 (3) ◽

pp. 180 ◽

Cited By ~ 10

Author(s):

RP Goswami ◽

S Das ◽

RP Goswami ◽

Y Ray ◽

M Rahman

Keyword(s):

Visceral Leishmaniasis ◽

Early Report ◽

Urine Samples ◽

Eastern India ◽

Non Invasive

Download Full-text

Detection of asymptomatic Leishmania infection in Bangladesh by novel antigen and antibody diagnostic tools shows strong association with PKDL patients

10.1101/2020.05.26.20113431 ◽

2020 ◽

Author(s):

Sophie Owen ◽

Faria Hossain ◽

Prakash Ghosh ◽

Rajashree Chowdhury ◽

Md. Sakhawat Hossain ◽

...

Keyword(s):

Visceral Leishmaniasis ◽

Quantitative Polymerase Chain Reaction ◽

Asymptomatic Infection ◽

Diagnostic Tools ◽

Leishmania Infection ◽

Active Infection ◽

Non Invasive ◽

Antigen Elisa ◽

Disease Reservoirs ◽

Index Cases

AbstractBackgroundAsymptomatic Leishmania infections outnumber clinical infections on the Indian sub-continent (ISC) where disease reservoirs are anthroponotic. Diagnostics which detect active asymptomatic infection, which are suitable for monitoring and surveillance, may be of benefit to the visceral leishmaniasis (VL) elimination campaign on the ISC.Methodology/Principal FindingsQuantitative polymerase chain reaction (qPCR), loop mediated isothermal amplification (LAMP), the direct agglutination test (DAT), and the Leishmania antigen ELISA were carried out on blood and urine samples collected from 720 household and neighbouring contacts of 276 VL and post kala-azar dermal leishmaniasis (PKDL) index cases, with no symptoms or history of VL and PKDL, in endemic regions of Bangladesh between September 2016 and March 2018. Of the 720 contacts of index cases, asymptomatic infection was detected in 69 (9.6%) participants by a combination of qPCR (1.0%), LAMP (2.1%), DAT (3.9%), and Leishmania antigen ELISA (3.3%). Only 1 (0.1%) participant was detected positive by all 4 diagnostic tests. Poor agreement between tests was calculated using Cohen’s kappa (k) statistics, however the Leishmania antigen ELISA and DAT in combination capture all participants positive by more than one test. We find strong evidence for association between the index case being a PKDL case (OR 1.94, p = 0.009), specifically macular PKDL (OR 2.12, p = 0.004) and being positive for at least one of the four tests.Conclusions/SignificanceLeishmania antigen ELISA detects active asymptomatic infection, requires a non-invasive sample, and therefore may be of benefit for monitoring transmission and surveillance in an elimination setting in combination with serology. Development of an antigen detection test in RDT format would be of benefit to the elimination campaign.Author summaryInfection with the parasite Leishmania donovani can lead to an asymptomatic infection with only around 5% of asymptomatics converting to visceral leishmaniasis the clinical manifestation of the infection. Serological assays detect anti-Leishmania antibodies and therefore cannot distinguish between past and active infection. Molecular assays and those which detect Leishmania antigens detect active infection. Since the signing of a memorandum of understanding in 2005, visceral leishmaniasis has been targeted for elimination in India, Nepal and Bangladesh. In an elimination setting such as Bangladesh, where disease reservoirs are anthroponotic, a relatively simple test such as the Leishmania antigen ELISA which requires a non-invasive urine sample, may be of benefit in combination with serology for surveillance and monitoring of Leishmania transmission. Development of an antigen test into a field compatible rapid diagnostic test would be of further benefit to the elimination campaign.

Download Full-text

926: A Pilot Study to Determine Local Oxygen Saturation of the Flaccid and Erect Penis in Men with and without Erectile Dysfunction Using a Non-Invasive Tissue Oximeter

The Journal of Urology ◽

10.1016/s0022-5347(18)33162-8 ◽

2006 ◽

Vol 175 (4S) ◽

pp. 300-300 ◽

Cited By ~ 1

Author(s):

Priya Padmanabhan ◽

Andrew R. McCullough

Keyword(s):

Erectile Dysfunction ◽

Pilot Study ◽

Oxygen Saturation ◽

Non Invasive ◽

Local Oxygen ◽

Erect Penis

Download Full-text

Non-Invasive Distinction of Non-Alcoholic Fatty Liver Disease using Urinary Volatile Organic Compound Analysis: Early Results

Journal of Gastrointestinal and Liver Diseases ◽

10.15403/jgld.2014.1121.242.ury ◽

2015 ◽

Vol 24 (2) ◽

pp. 197-201 ◽

Cited By ~ 10

Author(s):

Ramesh P. Arasaradnam ◽

Michael McFarlane ◽

Emma Daulton ◽

Erik Westenbrink ◽

Nicola O’Connell ◽

...

Keyword(s):

Liver Disease ◽

Pilot Study ◽

Fatty Liver ◽

Fatty Liver Disease ◽

Diagnostic Methods ◽

Screening Methods ◽

Alcoholic Fatty Liver ◽

Non Invasive ◽

Volatile Organic ◽

Alcoholic Fatty Liver Disease

Background & Aims: Non-Alcoholic Fatty Liver Disease (NAFLD) is the commonest cause of chronic liver disease in the western world. Current diagnostic methods including Fibroscan have limitations, thus there is a need for more robust non-invasive screening methods. The gut microbiome is altered in several gastrointestinal and hepatic disorders resulting in altered, unique gut fermentation patterns, detectable by analysis of volatile organic compounds (VOCs) in urine, breath and faeces. We performed a proof of principle pilot study to determine if progressive fatty liver disease produced an altered urinary VOC pattern; specifically NAFLD and Non-Alcoholic Steatohepatitis (NASH).Methods: 34 patients were recruited: 8 NASH cirrhotics (NASH-C); 7 non-cirrhotic NASH; 4 NAFLD and 15 controls. Urine was collected and stored frozen. For assay, the samples were defrosted and aliquoted into vials, which were heated to 40±0.1°C and the headspace analyzed by FAIMS (Field Asymmetric Ion Mobility Spectroscopy). A previously used data processing pipeline employing a Random Forrest classification algorithm and using a 10 fold cross validation method was applied.Results: Urinary VOC results demonstrated sensitivity of 0.58 (0.33 - 0.88), but specificity of 0.93 (0.68 - 1.00) and an Area Under Curve (AUC) 0.73 (0.55 -0.90) to distinguish between liver disease and controls. However, NASH/NASH-C was separated from the NAFLD/controls with a sensitivity of 0.73 (0.45 - 0.92), specificity of 0.79 (0.54 - 0.94) and AUC of 0.79 (0.64 - 0.95), respectively.Conclusions: This pilot study suggests that urinary VOCs detection may offer the potential for early non-invasive characterisation of liver disease using 'smell prints' to distinguish between NASH and NAFLD.

Download Full-text

Evaluation of qPCR on blood and skin microbiopsies, peripheral blood buffy coat smear, and urine antigen ELISA for diagnosis and test of cure for visceral leishmaniasis in HIV-coinfected patients in India: a prospective cohort study

BMJ Open ◽

10.1136/bmjopen-2020-042519 ◽

2021 ◽

Vol 11 (4) ◽

pp. e042519

Author(s):

Sophie I Owen ◽

Sakib Burza ◽

Shiril Kumar ◽

Neena Verma ◽

Raman Mahajan ◽

...

Keyword(s):

Bone Marrow ◽

Visceral Leishmaniasis ◽

Hiv Infection ◽

Peripheral Blood ◽

Medical Science ◽

Bone Marrow Aspiration ◽

Buffy Coat ◽

Tropical Medicine ◽

Urine Antigen ◽

Antigen Elisa

IntroductionHIV coinfection presents a challenge for diagnosis of visceral leishmaniasis (VL). Invasive splenic or bone marrow aspiration with microscopic visualisation of Leishmania parasites remains the gold standard for diagnosis of VL in HIV-coinfected patients. Furthermore, a test of cure by splenic or bone marrow aspiration is required as patients with VL-HIV infection are at a high risk of treatment failure. However, there remain financial, implementation and safety costs to these invasive techniques which severely limit their use under field conditions.Methods and analysisWe aim to evaluate blood and skin qPCR, peripheral blood buffy coat smear microscopy and urine antigen ELISA as non-invasive or minimally invasive alternatives for diagnosis and post-treatment test of cure for VL in HIV-coinfected patients in India, using a sample of 91 patients with parasitologically confirmed symptomatic VL-HIV infection.Ethics and disseminationEthical approval for this study has been granted by The Liverpool School of Tropical Medicine, The Institute of Tropical Medicine in Antwerp, the University of Antwerp and the Rajendra Memorial Research Institute of Medical Science in Patna. Any future publications will be published in open access journals.Trial registration numberCTRI/2019/03/017908.

Download Full-text

Does transcranial direct current stimulation affect selective visual attention in children with left-sided infantile hemiplegia? A randomized, controlled pilot study

Brain Impairment ◽

10.1017/brimp.2020.20 ◽

2020 ◽

pp. 1-13

Author(s):

Raed A. Alharbi ◽

Saleh A. Aloyuni ◽

Faizan Kashoo ◽

Mohamed I. Waly ◽

Harpreet Singh ◽

...

Keyword(s):

Pilot Study ◽

Direct Current ◽

Transcranial Direct Current Stimulation ◽

Repeated Measures ◽

Sham Group ◽

Time Points ◽

Selective Visual Attention ◽

Subject Factor ◽

And Control

Abstract Objective: Infantile hemiplegia due to brain injury is associated with poor attention span, which critically affects the learning and acquisition of new skills, especially among children with left-sided infantile hemiplegia (LSIH). This study aimed to improve the selective visual attention (SVA) of children with LSIH through transcranial direct current stimulation (tDCS). Methods: A total of 15 children participated in this randomized, double-blinded, pilot study; of them, 10 experienced LSIH, and the remaining 5 were healthy age-matched controls. All the children performed the Computerized Stroop Color-Word Test (CSCWT) at baseline, during the 5th and 10th treatment sessions, and at follow-up. The experimental (n = 5) and control groups (n = 5) received tDCS, while the sham group (n = 5) received placebo tDCS. All three groups received cognitive training on alternate days, for 3 weeks, with the aim to improve SVA. Results: Two-way repeated measures analysis of variance (ANOVA) showed a statistically significant change in the mean scores of CSCWT between time points (baseline, 5th and 10th sessions, and follow-up) within-subject factor, group (experimental, sham) between-subject factor and interaction (time points X group) (p < 0.005). Furthermore, a one-way repeated measures ANOVA showed significant differences between time point (p < 0.005) for the experimental and control group but not the sham group. Conclusion: These pilot results suggest that future research should be conducted with adequate samples to enable conclusions to be drawn.

Download Full-text

A non-invasive method to generate induced pluripotent stem cells from primate urine

Scientific Reports ◽

10.1038/s41598-021-82883-0 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Johanna Geuder ◽

Lucas E. Wange ◽

Aleksandar Janjic ◽

Jessica Radmer ◽

Philipp Janssen ◽

...

Keyword(s):

Stem Cells ◽

Induced Pluripotent Stem Cells ◽

Pluripotent Stem Cells ◽

Sendai Virus ◽

Cell Types ◽

Urine Samples ◽

Comparative Approach ◽

Non Invasive ◽

Human Ipscs ◽

Induced Pluripotent

AbstractComparing the molecular and cellular properties among primates is crucial to better understand human evolution and biology. However, it is difficult or ethically impossible to collect matched tissues from many primates, especially during development. An alternative is to model different cell types and their development using induced pluripotent stem cells (iPSCs). These can be generated from many tissue sources, but non-invasive sampling would decisively broaden the spectrum of non-human primates that can be investigated. Here, we report the generation of primate iPSCs from urine samples. We first validate and optimize the procedure using human urine samples and show that suspension- Sendai Virus transduction of reprogramming factors into urinary cells efficiently generates integration-free iPSCs, which maintain their pluripotency under feeder-free culture conditions. We demonstrate that this method is also applicable to gorilla and orangutan urinary cells isolated from a non-sterile zoo floor. We characterize the urinary cells, iPSCs and derived neural progenitor cells using karyotyping, immunohistochemistry, differentiation assays and RNA-sequencing. We show that the urine-derived human iPSCs are indistinguishable from well characterized PBMC-derived human iPSCs and that the gorilla and orangutan iPSCs are well comparable to the human iPSCs. In summary, this study introduces a novel and efficient approach to non-invasively generate iPSCs from primate urine. This will extend the zoo of species available for a comparative approach to molecular and cellular phenotypes.

Download Full-text