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 ◽  
...  

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.

2020 ◽  
Vol 58 (11) ◽  
Author(s):  
Nicky Didwania ◽  
Sarfaraz Ahmad Ejazi ◽  
Rudra Chhajer ◽  
Abdus Sabur ◽  
Saumyabrata Mazumder ◽  
...  

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.


2002 ◽  
Vol 9 (4) ◽  
pp. 789-794 ◽  
Author(s):  
Mohammad Zahidul Islam ◽  
Makoto Itoh ◽  
S. M. Shamsuzzaman ◽  
Rusella Mirza ◽  
Farzana Matin ◽  
...  

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.


PPAR Research ◽  
2012 ◽  
Vol 2012 ◽  
pp. 1-11 ◽  
Author(s):  
Marion M. Chan ◽  
Nagasuresh Adapala ◽  
Cui Chen

Infection is the outcome of a contest between a pathogen and its host. In the disease leishmaniasis, the causative protozoan parasites are harbored inside the macrophages.Leishmaniaspecies adapt strategies to make the infection chronic, keeping a balance between their own and the host's defense so as to establish an environment that is favorable for survival and propagation. Activation of peroxisome proliferator-activated receptor (PPAR) is one of the tactics used. This ligand-activated nuclear factor curbs inflammation to protect the host from excessive injuries by setting a limit to its destructive force. In this paper, we report the interaction of host PPARs and the pathogen for visceral leishmaniasis,Leishmania donovani,in vivoandin vitro. PPAR expression is induced by parasitic infection. Leishmanial activation of PPARγpromotes survival, whereas blockade of PPARγfacilitates removal of the parasite. Thus,Leishmaniaparasites harness PPARγto increase infectivity.


Immunobiology ◽  
2021 ◽  
Vol 226 (2) ◽  
pp. 152057
Author(s):  
Deepak Kumar Goyal ◽  
Poonam Keshav ◽  
Sukhbir Kaur

2021 ◽  
Vol 9 ◽  
pp. 205031212110367
Author(s):  
Berhanu Tarekegn ◽  
Ayanaw Tamene

Background: Visceral leishmaniasis is a vector-borne disease caused by Leishmania donovani transmitted by sand fly species. It is the third most common vector-borne disease globally. Visceral leishmaniasis is endemic in Ethiopia with an estimated annual incidence ranging from 3700 to 7400 cases. This research aimed to assess the clinical presentations and laboratory profiles of visceral leishmaniasis for early diagnosis and timely initiation of management. Objective: To describe the clinical and laboratory manifestation and diagnostic modalities of visceral leishmaniasis among adult patients admitted to Felege Hiwot Hospital, from 1 September 2016 to 30 August 2019. Method: Institution-based retrospective cross-sectional study was conducted among 141 patients admitted to Felege Hiwot Hospital from 1 September 2016 to 30 August 2019. Descriptive statistics were used to describe the clinical presentation and laboratory profiles of patients with visceral leishmaniasis. Results: Among a total of 141 enrolled patients in the study, males were affected 13-fold. Most of them were travelers to endemic areas during the winter season for labor work. The mean duration of illness was 48 days. Common symptoms were fever (96.5%), weightless (82.5%), jaundice (18.4%), vomiting/diarrhea (13.5%), and bleeding episodes (11.3%). Splenomegaly was seen in 98.6%, ascites in 35.5%, and lymphadenopathy in 9.9%. Lymphadenopathy was seen significantly in HIV patients (40%). Anemia was seen in 95%, thrombocytopenia in 90.2%, leukopenia in 86.4%, and pancytopenia in 79.4%. Half of the patients had coinfection. Neutropenic sepsis was seen in 21.3%. The diagnosis was made by tissue aspiration in 65% of patients. Conclusion: The majority of patients who were diagnosed to have visceral leishmaniasis were young male adults who traveled to the endemic areas seasonally. Fever and splenomegaly were seen as the commonest clinical presentation. Lymphadenopathy occurred in high frequency among HIV co-infected patients. Anemia was the commonest hematologic finding.


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