scholarly journals Evaluation of Cysteine Protease C of Leishmania donovani in Comparison with Glycoprotein 63 and Elongation Factor 1α for Diagnosis of Human Visceral Leishmaniasis and for Posttreatment Follow-Up Response

2020 ◽  
Vol 58 (11) ◽  
Author(s):  
Nicky Didwania ◽  
Sarfaraz Ahmad Ejazi ◽  
Rudra Chhajer ◽  
Abdus Sabur ◽  
Saumyabrata Mazumder ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Human Serum ◽  
Cysteine Protease ◽  
Leishmania Donovani ◽  
Elongation Factor ◽  
Urine Samples ◽  
Healthy Controls ◽  
Elongation Factor 1Α ◽  
Antibody Levels ◽  
Serum And Urine

ABSTRACT Visceral leishmaniasis (VL) is a threat in many developing countries. Much effort has been put to eliminating this disease, for which serodiagnosis remains the mainstay for VL control programs. New and improved antigens as diagnostic candidates are required, though, as the available antigens fail to demonstrate equal optimum performance in all areas of endemicity. Moreover, these diagnoses are dependent on invasive serum sampling. In the current study, we cloned and expressed Leishmania donovani cysteine protease C (CPC) and evaluated its diagnostic and test-of-cure possibilities by detecting the antibody levels in human serum and urine through ELISA and immunoblot assays. Two immunodominant antigens, recombinant glycoprotein 63 (GP63) and elongation factor 1α (EF1α), identified earlier by our group, were also assessed by employing human serum and urine samples. Of these three antigens in ELISAs, CPC demonstrated the highest sensitivities of 98.15% and 96% positive testing in serum and urine of VL patients, respectively. Moreover, CPC yielded 100% specificity with serum and urine of nonendemic healthy controls compared to GP63 and EF1α. Urine samples were found to be more specific than serum for distinguishing endemic healthy controls and other diseases by means of all three antigens. In all cases, CPC gave the most promising results. Unlike serum, urine tests demonstrated a significant decrease in antibody levels for CPC, GP63, and EF1α after 6 months of treatment. The diagnostic and test-of-cure performances of CPC in the immunoblot assay were found to be better than those of GP63 and EF1α. In conclusion, CPC, followed by GP63 and EF1α, may be utilized as candidates for diagnosis of VL and to assess treatment response.

scholarly journals Diagnosis of Visceral Leishmaniasis by Enzyme-Linked Immunosorbent Assay Using Urine Samples

2002 ◽  
Vol 9 (4) ◽  
pp. 789-794 ◽  
Author(s):  
Mohammad Zahidul Islam ◽  
Makoto Itoh ◽  
S. M. Shamsuzzaman ◽  
Rusella Mirza ◽  
Farzana Matin ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Sensitivity And Specificity ◽  
Immunosorbent Assay ◽  
Leishmania Donovani ◽  
Laboratory Diagnosis ◽  
Urine Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Controls ◽  
Serum Samples ◽  
Crude Antigen

ABSTRACT A diagnostic method has been developed to detect anti-Leishmania donovani immunoglobulin G (IgG) in urine by enzyme-linked immunosorbent assay (ELISA). In measuring anti-L. donovani IgG, IgA, and IgM in urine, the method performed best in the detection of IgG. The sensitivity and specificity of the assay were determined with panels of urine samples from 62 visceral leishmaniasis (VL) patients, 59 healthy controls from areas of endemicity, 53 healthy controls from areas of nonendemicity, 59 malaria patients, 13 tuberculosis patients, 23 cutaneous leishmaniasis patients, and 7 patients with other diseases. Using L. donovani promastigote crude antigen, the test had 93.5% sensitivity (58 positives of 62 VL patient samples) and 89.3% specificity (191 negatives of 214 non-VL patient samples). The ELISA with acetone-treated L. donovani promastigote antigen raised the sensitivity and specificity to 95.0 and 95.3%, respectively. Western blot analysis revealed that most of the samples that cross-reacted with crude antigen in ELISA did not recognize any antigenic component of L. donovani crude antigen. We also checked 40 serum samples from the same group of VL patients for anti-L. donovani IgG and got 90.0% sensitivity with both crude and acetone-treated antigens. As collection of urine is much easier than collection of serum, the detection of anti-L. donovani IgG in urine with acetone-treated antigen will be useful in epidemiological studies. It could be an adjunct of laboratory diagnosis.

scholarly journals Development and Clinical Evaluation of Serum and Urine-Based Lateral Flow Tests for Diagnosis of Human Visceral Leishmaniasis

2021 ◽  
Vol 9 (7) ◽  
pp. 1369
Author(s):  
Sarfaraz Ahmad Ejazi ◽  
Somsubhra Thakur Choudhury ◽  
Anirban Bhattacharyya ◽  
Mohd Kamran ◽  
Krishna Pandey ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Donovani ◽  
Parasitic Infection ◽  
Lateral Flow ◽  
Urine Samples ◽  
Experimental Conditions ◽  
Long Time ◽  
Lateral Flow Tests ◽  
Imagej Software ◽  
Serum And Urine

Visceral leishmaniasis (VL), a fatal parasitic infection, is categorized as being neglected among tropical diseases. The use of conventional tissue aspiration for diagnosis is not possible in every setting. The immunochromatography-based lateral flow assay (LFA) has attracted attention for a long time due to its ability to give results within a few minutes, mainly in resource-poor settings. In the present study, we optimized and developed the LFA to detect anti-Leishmania antibodies for VL diagnosis. The performance of the developed test was evaluated with serum and urine samples of Indian VL patients and Brazilian sera. The new test exploits well-studied and highly-sensitive purified antigens, LAg isolated from Leishmania donovani promastigotes and protein G conjugated colloidal-gold as a signal reporter. The intensity of the bands depicting the antigen–antibody complex was optimized under different experimental conditions and quantitatively analyzed by the ImageJ software. For the diagnosis of human VL in India, LFA was found to be 96.49% sensitive and 95% specific with serum, and 95.12% sensitive and 96.36% specific with urine samples, respectively. The sensitivity and specificity of LFA were 88.57% and 94.73%, respectively, for the diagnosis of Brazilian VL using patients’ sera infected with Leishmania infantum. LFA is rapid and simple to apply, suitable for field usage where results can be interpreted visually and particularly sensitive and specific in the diagnosis of human VL. Serum and urine LFA may improve diagnostic outcomes and could be an alternative for VL diagnosis in settings where tissue aspiration is difficult to perform.

scholarly journals Per- and polyfluoroalkyl substances in human serum and urine samples from a residentially exposed community

2017 ◽  
Vol 106 ◽  
pp. 135-143 ◽  
Author(s):  
Rachel Rogers Worley ◽  
Susan McAfee Moore ◽  
Bruce C. Tierney ◽  
Xiaoyun Ye ◽  
Antonia M. Calafat ◽  
...  
Keyword(s):  
Human Serum ◽  
Urine Samples ◽  
Polyfluoroalkyl Substances ◽  
Serum And Urine ◽  
Serum And Urine Samples

scholarly journals Antibody and cytokine levels in visceral leishmaniasis patients with varied parasitemia before, during, and after treatment in patients admitted to Arba Minch General Hospital, southern Ethiopia

2021 ◽  
Vol 15 (8) ◽  
pp. e0009632
Author(s):  
Dagimawie Tadesse ◽  
Alemseged Abdissa ◽  
Mekidim Mekonnen ◽  
Tariku Belay ◽  
Asrat Hailu
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Donovani ◽  
Serum Levels ◽  
High Serum ◽  
Eastern Africa ◽  
Southern Ethiopia ◽  
Ifn Γ ◽  
A Prospective Study ◽  
Antibody Levels ◽  
Tgf Β1

Background Visceral leishmaniasis is a disease caused by disseminated Leishmania donovani infection which affects almost half a million people annually. Most of the patients are reported from the Indian sub-continent, Eastern Africa and Brazil. In this study, we aimed to determine the levels of antibodies and cytokines in visceral leishmaniasis patients and to examine associations of parasitemia with the clinical states of patients. A prospective study was carried out, enrolling a total of 48 active VL patients who were evaluated before, during different time points and, three months after treatment. Serum cytokine concentrations, antibody levels, parasitemia, laboratory (hematologic and biochemical) measurements, and clinical parameters were assessed. Results Counts of WBC and platelets, and measurements of hemoglobin (Hb) increased during treatment (P ≤ 0.05). Elevated levels of circulating IL-10, IFN-γ, and TGF-β1 were measured before treatment. The observed increase in serum IL-10 remarkably declined within 7 days after the start of treatment. Anti-leishmanial antibody index (AI) was high in all VL patients irrespective of spleen aspirate parasite grade before treatment and at different times during treatment. However, a significant (P ≤ 0.05) decrease of AI was observed 120 days post-treatment. IL-2 serum levels were below the detection limit at all sampling points. Conclusions The present results suggest that IL-10, IFN-γ, and TGF-β1 can be used as markers of active visceral leishmaniasis. In addition, measuring circulating cytokines concentrations, particularly IL-10, in combination with other clinical evaluations, could be used as criteria for the cure. The observation that a high serum concentration of IFN-gamma at baseline was associated with low parasitemia deserves further investigations.

scholarly journals A Simple RP-HPLC Method for the Determination of Omeprazole in Human Serum and Urine: Validation and Application in Pharmacokinetic Study

1970 ◽  
Vol 8 (2) ◽  
pp. 123-130 ◽  
Author(s):  
Kamrun Nahar ◽  
Jafreen Jamal Joti ◽  
Md Ashik Ullah ◽  
Ahasanul Hasan ◽  
Mohammad Abul Kalam Azad ◽  
...  
Keyword(s):  
Human Serum ◽  
Hplc Method ◽  
Stability Study ◽  
Urine Samples ◽  
Dihydrogen Phosphate ◽  
Serum Samples ◽  
Rp Hplc ◽  
Limit Of Quantitation ◽  
Serum And Urine ◽  
Serum And Urine Samples

A Simple RP-HPLC method with UV detection has been validated to determine omeprazole concentrations in human serum and urine samples. The mobile phase consisted of a mixture of potassium dihydrogen phosphate buffer (pH 7.2 ± 0.05; 0.2 M) and acetonitrile (70:30, v/v), pumped at a flow rate of 1.0 ml/min through the C-8 column at room temperature. Peaks were monitored by UV absorbance at 302 nm at a sensitivity of 0.0001. The developed method was selective and linear for omeprazole concentrations ranging between 5 to 1000ng/ml for serum samples and 1 to 100μg/ml for urine samples. The recovery of omeprazole ranged from 95.68 to 99% and 95.54 to 99.8% for the serum and urine samples respectively. The limit of quantitation (LOQ) of omeprazole was 5 ng/ml. The intraday accuracy ranged from 93.54 to 104.38% and 100.55 to 103.48% for the serum and urine respectively. The interday accuracy varied from 97.61 to 113.95% and 97.42 to 109.97% for the serum and urine respectively. For the LOQ, good values of precision (6.03 and 10.13% for intraday and interday, respectively) were also obtained. Acceptable results were obtained during stability study. This method proved to be simple, accurate and precise for pharmacokinetic and bioequivalence studies of omeprazole. Key words: Omeprazole; RP-HPLC; Method validation. DOI: 10.3329/dujps.v8i2.6026 Dhaka Univ. J. Pharm. Sci. 8(2): 123-130, 2009 (December)

scholarly journals Electrochemical sensor formed from poly(3,4-ethylenedioxyselenophene) and nitrogen-doped graphene composite for dopamine detection

RSC Advances ◽  
2021 ◽  
Vol 11 (59) ◽  
pp. 37544-37551
Author(s):  
Aygul Kadir ◽  
Ruxangul Jamal ◽  
Tursun Abdiryim ◽  
Nurbiya Sawut ◽  
Yuzhu Che ◽  
...  
Keyword(s):  
Human Serum ◽  
Electrochemical Sensor ◽  
Urine Samples ◽  
Nitrogen Doped ◽  
Graphene Composite ◽  
Dopamine Detection ◽  
Nitrogen Doped Graphene ◽  
Doped Graphene ◽  
Serum And Urine ◽  
Serum And Urine Samples

An electrochemical sensor for dopamine detection has been fabricated using a composite of PEDOS and N-Gr. The results of actual samples showed that the composite of PEDOS/N-Gr has excellent recovery (95.40–100.14%) for human serum and urine samples.

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