scholarly journals Detection of Immunoglobulin G1 Against rK39 Improves Monitoring of Treatment Outcomes in Visceral Leishmaniasis

2018 ◽  
Vol 69 (7) ◽  
pp. 1130-1135 ◽  
Author(s):  
Guy Mollett ◽  
Bruno C Bremer Hinckel ◽  
Tapan Bhattacharyya ◽  
Tegwen Marlais ◽  
Om Prakash Singh ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Treatment Outcomes ◽  
Strong Evidence ◽  
Diagnostic Tests ◽  
Leishmania Donovani ◽  
Clinical Status ◽  
Post Treatment ◽  
Serum Samples ◽  
Significant Difference ◽  
Early Case

Abstract Background Visceral leishmaniasis (VL), caused by the Leishmania donovani complex, is a fatal, neglected tropical disease that is targeted for elimination in India, Nepal, and Bangladesh. Improved diagnostic tests are required for early case detection and for monitoring the outcomes of treatments. Previous investigations using Leishmania lysate antigen demonstrated that the immunoglobulin (Ig) G1 response is a potential indicator of a patient’s clinical status after chemotherapy. Methods IgG1 or IgG enzyme-linked immunosorbent assays (ELISAs) with rK39 or lysate antigens and novel IgG1 rK39 rapid diagnostic tests (RDTs) were assessed with Indian VL serum samples from the following clinical groups: paired pre- and postchemotherapy (deemed cured); relapsed; other infectious diseases; and endemic, healthy controls. Results With paired pre- and post-treatment samples (n = 37 pairs), ELISAs with rK39- and IgG1-specific conjugates gave a far more discriminative decrease in post-treatment antibody responses when compared to IgG (P < .0001). Novel IgG1 rK39 RDTs provided strong evidence for decreased IgG1 responses in patients who had successful treatment (P < .0001). Furthermore, both IgG1 rK39 RDTs (n = 38) and ELISAs showed a highly significant difference in test outcomes between cured patients and those who relapsed (n = 23; P < .0001). RDTs were more sensitive than corresponding ELISAs. Conclusions We present strong evidence for the use of IgG1 in monitoring treatment outcomes in VL, and the first use of an IgG1-based RDT using the rK39 antigen for the discrimination of post-treatment cure versus relapse in VL. Such an RDT may have a significant role in monitoring patients and in targeted control and elimination of this devastating disease.

scholarly journals Phospholipase A2 (PLA2) as an Early Indicator of Envenomation in Australian Elapid Snakebites (ASP-27)

Biomedicines ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 459
Author(s):  
Geoffrey K. Isbister ◽  
Nandita Mirajkar ◽  
Kellie Fakes ◽  
Simon G. A. Brown ◽  
Punnam Chander Veerati
Keyword(s):  
Characteristic Curve ◽  
Diagnostic Value ◽  
Pla2 Activity ◽  
Serum Samples ◽  
Case Identification ◽  
Chemical Company ◽  
Significant Difference ◽  
Ann Arbor ◽  
Early Case ◽  
Cayman Chemical

Early diagnosis of snake envenomation is essential, especially neurotoxicity and myotoxicity. We investigated the diagnostic value of serum phospholipase (PLA2) in Australian snakebites. In total, 115 envenomated and 80 non-envenomated patients were recruited over 2 years, in which an early blood sample was available pre-antivenom. Serum samples were analyzed for secretory PLA2 activity using a Cayman sPLA2 assay kit (#765001 Cayman Chemical Company, Ann Arbor MI, USA). Venom concentrations were measured for snake identification using venom-specific enzyme immunoassay. The most common snakes were Pseudonaja spp. (33), Notechis scutatus (24), Pseudechis porphyriacus (19) and Tropidechis carinatus (17). There was a significant difference in median PLA2 activity between non-envenomated (9 nmol/min/mL; IQR: 7–11) and envenomated patients (19 nmol/min/mL; IQR: 10–66, p < 0.0001) but Pseudonaja spp. were not different to non-envenomated. There was a significant correlation between venom concentrations and PLA2 activity (r = 0.71; p < 0.0001). PLA2 activity was predictive for envenomation; area under the receiver-operating-characteristic curve (AUC-ROC), 0.79 (95% confidence intervals [95%CI]: 0.72–0.85), which improved with brown snakes excluded, AUC-ROC, 0.88 (95%CI: 0.82–0.94). A cut-point of 16 nmol/min/mL gives a sensitivity of 72% and specificity of 100% for Australian snakes, excluding Pseudonaja. PLA2 activity was a good early predictor of envenomation in most Australian elapid bites. A bedside PLA2 activity test has potential utility for early case identification but may not be useful for excluding envenomation.

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 Performance of an indigenous β-mercaptoethanol-modified antigen in comparison with a commercial reference in direct agglutination test for detection of canine visceral leishmaniasis

2014 ◽  
Vol 63 (1) ◽  
pp. 106-110
Author(s):  
Saul José Semião-Santos ◽  
Laura Barral Veloso ◽  
Paulo Paes de Andrade ◽  
Marcia Almeida de Melo ◽  
Luis Miguel Lourenço Martins ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Infantum ◽  
Leishmania Donovani ◽  
Agglutination Test ◽  
Canine Visceral Leishmaniasis ◽  
Direct Agglutination Test ◽  
Serum Samples

We compared the performance of a locally produced β-mercaptoethanol-modified promastigote antigen (β-ME-Ag) of an indigenous Leishmania infantum strain against that of a trypsinized Leishmania donovani reference (REF-Ag) in the direct agglutination test (DAT) for detection of canine visceral leishmaniasis (CVL). One hundred and fifty-one serum samples collected from dogs belonging to four groups with different conditions were included. At a DAT titre of 1 : 320, statistically determined as optimal cut-off value for β-ME-Ag, and 1 : 160 for REF-Ag, a sensitivity and a specificity of 100 % were estimated for β-ME-Ag in comparison with 96.6 % and 100 %, respectively, for REF-Ag. Overall, levels of agglutination titres recorded for the two antigens were highly concordant (Cohen’s κ = 0.879) in both the CVL and non-CVL groups. Based on current results, and ease experienced in processing the antigen and reading the test outcome, we recommend incorporation of β-ME-Ag in DAT for confirmation or exclusion of suspected CVL in dogs.

scholarly journals L. (L.) chagasi in aids and visceral leishmaniasis (kala-azar) co-infection

2008 ◽  
Vol 50 (4) ◽  
pp. 251-254 ◽  
Author(s):  
Ana Maria Roselino ◽  
Maria Fernanda Chociay ◽  
Roberto Silva Costa ◽  
Alcyone Artioli Machado ◽  
José Fernando de Castro Figueiredo
Keyword(s):  
Visceral Leishmaniasis ◽  
Skin Biopsy ◽  
Leishmania Donovani ◽  
Skin Lesions ◽  
Post Treatment ◽  
Kala Azar ◽  
Tegumentary Leishmaniasis

Concomitant skin lesions in visceral leishmaniasis (VL) or kala-azar are rare, being more common the description of post-kala-azar dermal leishmaniasis occurring post treatment of kala-azar. Skin lesions caused by Leishmania donovani are frequently seen in the aids-VL co-infection. In Brazil cutaneous or mucosal forms of tegumentary leishmaniasis concomitant with aids are more commonly registered. Here we present a case of aids-VL co-infection, with unusual cutaneous and digestive compromising attributed to L. (L.) chagasi, with special attention to ecthymatous aspect of the lesion, allied to the absence of parasite on the histological skin biopsy.

scholarly journals Comparing the impact of genotypic based diagnostic algorithm on time to treatment initiation and treatment outcomes among drug-resistant tuberculosis patients in Amhara region, Ethiopia

PLoS ONE ◽  
2021 ◽  
Vol 16 (2) ◽  
pp. e0246938
Author(s):  
Getahun Molla Kassa ◽  
Mehari Woldemariam Merid ◽  
Atalay Goshu Muluneh ◽  
Haileab Fekadu Wolde
Keyword(s):  
Treatment Outcomes ◽  
Median Time ◽  
Diagnostic Tests ◽  
Treatment Initiation ◽  
Diagnostic Tools ◽  
Care Seeking ◽  
Solid Culture ◽  
Significant Difference ◽  
Probe Assay ◽  
The Impact

Background To end Tuberculosis (TB) by 2030, early detection and timely treatment of Drug-Resistant Tuberculosis (DR-TB) is vital. The role of rapid, accurate, and sensitive DR-TB diagnostic tool is indispensable to accelerate the TB control program. There are evidence breaks in the time difference and its effect on treatment outcomes among different DR-TB diagnostic tools in Ethiopia. This article aimed to compare the different DR-TB diagnostic tools with time pointers and evaluate their effect on the treatment outcomes. Method We performed a retrospective chart review of 574 DR-TB patients from September 2010 to December 2017 to compare the impact of molecular DR-TB diagnostic tests (Xpert MTB/RIF, Line Probe Assay (LPA), and solid culture-based Drug Susceptibility Testing (DST)) on time to diagnosis, treatment initiation, and treatment Outcomes. Kruskual-Wallis test was employed to assess the presence of a significant difference in median time among the DR-TB diagnostic tests. Chi-Square and Fisher exact tests were used to test the presence of relations between treatment outcome and diagnostic tests. Result The data of 574 DR-TB patients were included in the analysis. From these, 321, 173, and 80 patients were diagnosed using Xpert MTB/RIF, Line Probe Assay (LPA), and solid culture-based DST, respectively. The median time in a day with (Interquartile range (IQR)) for Xpert MTB/RIF, LPA, and solid culture-based DST was from a first care-seeking visit to diagnosis: 2(0, 9), 4(1, 55), and 70(18, 182), from diagnosis to treatment initiation: 3(1, 8), 33(4, 76), and 44(9, 145), and from a first care-seeking visit to treatment initiation: 4(1, 11), 3(1, 12) and 76(3.75, 191) respectively. The shorter median time was observed in the Xpert MTB/RIF followed by the LPA, and this was statistically significant with a p-value <0.001. There was no statistically significant difference concerning treatment outcomes among the three DST tests. Conclusion Xpert MTB/RIF can mitigate the transmission of DR-TB significantly via quick diagnosis and treatment initiation followed by LPA as equating to the solid culture base DST, particularly in smear-positive patients. However, we didn’t see a statistically significant impact in terms of treatment outcomes. Xpert MTB/RIF can be used as the first test to diagnose DR-TB by further complimenting solid culture base DST to grasp the drug-resistance profile.

scholarly journals Immunoblot Analysis of the Humoral Immune Response to Leishmania donovani Polypeptides in Cases of Human Visceral Leishmaniasis: Its Usefulness in Prognosis

2002 ◽  
Vol 9 (5) ◽  
pp. 1119-1123 ◽  
Author(s):  
Promod Kumar ◽  
Kalpana Pai ◽  
Kiran Tripathi ◽  
H. P. Pandey ◽  
Shyam Sundar
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Donovani ◽  
Immunoblot Analysis ◽  
Specific Pattern ◽  
Sensitive Test ◽  
Soluble Extract ◽  
Serum Samples ◽  
Humoral Immune ◽  
Human Visceral Leishmaniasis ◽  
Sodium Antimony Gluconate

ABSTRACT Sera from Indian patients with parasitologically confirmed visceral leishmaniasis were studied by immunoblot analysis in order to identify a specific pattern for Leishmania infection. A soluble extract of Leishmania donovani was used as antigen. At diagnosis the sera from patients with visceral leishmaniasis specifically recognized fractions represented by bands of 201 kDa (50% of serum samples), 193 kDa (60%), 147 kDa (50%), 120 kDa (60%), 100 kDa (50%), 80 kDa (80%), 70 kDa (70%), 65 kDa (100%), 50 kDa (50%), 36 kDa (50%), 20 kDa (70%), and 18 kDa (50%). The 65-kDa band, common to all patients infected with Leishmania parasites, was found at the time of diagnosis. However, the immunoblot pattern changed after patients were treated and cured with sodium antimony gluconate (SAG; n =10) or miltefosine (n =10), as was evident from blots of sera obtained pretreatment and at 1, 3, and 6 months posttreatment. At 6 months posttreatment, immunoblots of sera from patients on the SAG regimen showed the disappearance of all bands except the 70-kDa band. Similarly, sera from those on the miltefosine regimen showed the disappearance of all bands except the 65- and 70-kDa bands. This study shows that Western blot analysis is a sensitive test for detection of anti-Leishmania antibodies. Moreover, the persistence of reactivity with the 65- and 70-kDa bands in the sera of all groups shows its promise as a diagnostic and prognostic tool.

scholarly journals Emergent canine visceral leishmaniasis in Argentina: Comparative diagnostics and relevance to proliferation of human disease

2021 ◽  
Vol 15 (7) ◽  
pp. e0009552
Author(s):  
Kyoko Fujisawa ◽  
Charlotte Silcott-Niles ◽  
Poppy Simonson ◽  
Daniela Lamattina ◽  
Cristian A. Humeres ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Diagnostic Tests ◽  
Point Of Care ◽  
Clinical Status ◽  
Clinical Signs ◽  
Cross Reactivity ◽  
Buffy Coat ◽  
Mucocutaneous Leishmaniasis ◽  
Antibody Titres ◽  
Skin Samples

Background Visceral leishmaniasis (VL) is a zoonotic protozoal vector-borne disease that is a major public health challenge. In Argentina, canine (CVL) and human visceral leishmaniasis (HVL) have recently emerged. There is a lack of standardised diagnostic tests for CVL, which hinders control of CVL and HVL. Methodology/Principal findings Sampling was carried out in Puerto Iguazú, Argentina, comprising 190 asymptomatic, oligosymptomatic and polysymptomatic dogs. The following diagnostics were applied: microscopy of lymph node aspirate (LNA); three immunochromatographic rapid diagnostic tests (RDTs), prototype rK28-ICT, rK39-ICT (both Coris BioConcept), commercial rK39 (InBios); ELISA for IgG, IgG1 and IgG2, against rK28, rK39 or crude lysate antigen. DNA detection and analysis, with 30 dogs, was of the ITS1 region using skin samples, and loop-mediated isothermal amplification (LAMP; Eiken Loopamp) of buffy coat, skin scrape or LNA. 15.4% of dogs were positive by LNA microscopy. The rK28 RDT had higher seropositivity rate (61%) than either a prototype rK39 RDT (31.4%) or commercial rK39 RDT (18.8%), without cross-reactivity with six other pathogens. IgG anti-rK39 ELISA antibody titres, but not IgG2, were positively correlated with number of clinical signs. LAMP with LNA had a higher positivity rate than PCR; buffy coat sampling was more sensitive than skin scrape. ITS1 confirmed Leishmania (Leishmania) infantum as the agent of CVL. Leishmania (Viannia) spp. was detected in skin samples from two dogs, compatible with Leishmania (Viannia) braziliensis. Conclusions/Significance Seroprevalence confirmed rapid increase in CVL in Puerto Iguazú. The rK28 RDT test potentially has great value for improved point-of-care diagnosis. Given cost reduction and accessibility, commercial LAMP may be applicable to buffy coat. RDT biomarkers of CVL clinical status are required to combat spread of CVL and HVL. The presence of Viannia, perhaps as an agent of human mucocutaneous leishmaniasis (MCL), highlights the need for vigilance and surveillance.

scholarly journals Phase II validation study of the rK39 ELISA prototype for the diagnosis of canine visceral leishmaniasis in Brazil

2021 ◽  
Vol 37 (3) ◽  
Author(s):  
Fernanda Alvarenga Cardoso Medeiros ◽  
Job Alves de Souza Filho ◽  
José Ronaldo Barbosa ◽  
Lucas Edel Donato ◽  
Fabiano Borges Figueiredo ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Validation Study ◽  
Diagnostic Performance ◽  
Clinical Status ◽  
Control Measures ◽  
Canine Visceral Leishmaniasis ◽  
Reference Standard ◽  
Serum Samples ◽  
Transmission Cycle ◽  
Healthy Dogs

Abstract: Dogs are the main reservoirs in the domestic transmission cycle of visceral leishmaniasis, and the diagnosis is essential for the effectiveness of the control measures recommended by the Brazilian Ministry of Health. We assessed the diagnostic performance of the ELISA-Vetlisa/BIOCLIN prototype with serum samples from 200 dogs, in triplicate, including symptomatic, oligosymptomatic, asymptomatic, and healthy dogs, originated by two distinct panels (A and B) characterized by parasitological tests as the reference standard. In this study, the prototype kit showed a 99% sensitivity (95%CI: 94.5-100.0) and a 100% specificity (95%CI: 96.4-100.0). The sensitivity of the prototype kit did not vary significantly with the clinical status of the dogs. Considering the final result classification (positive or negative), agreement between the results of repeated tests was almost perfect (kappa = 0.99; 95%CI: 0.98-1.00). ELISA-Vetlisa/BIOCLIN is a promising option for the serological diagnosis of canine visceral leishmaniasis in Brazil.

Effects of X-Ray Exposure on Some CMI Regulatory Cytokines in Technicians Serum Samples

2018 ◽  
Vol 9 (4) ◽  
Author(s):  
Weam S Al-Hamadany
Keyword(s):  
Clinical Status ◽  
Cell Mediated Immunity ◽  
Normal Range ◽  
Serum Samples ◽  
X Ray ◽  
Immunosuppressive Factor ◽  
Elisa Technique ◽  
Immunological Process ◽  
Field Workers ◽  
Tomography Scan

Ionizing radiation considered as an immunosuppressive factor upon over dose of exposure. Radiation field workers usually following a periodic checkup to monitor changes in their clinical status. Cell Mediated Immunity (CMI) has an important cytokines that regulate this specific immunological process. This study estimated the Interleukins (IL-2, IL-12 and IL-18) levels in serum samples using ELISA technique. Serum samples were collected from X-ray Technicians (Radiography, Fluoroscopy and Computed Tomography Scan Technicians) working in AL-Muthanna governorate hospitals. A total of (60) technicians and (30) control were involved in this research. Results showed significant decrease in IL-2 levels and increase in IL-18 levels in test groups (technicians) as compared with controls. While, IL-12 levels did not show a difference; all obtained values were within normal range. Overdose of X-ray exposure caused CMI suppression via disturbing the levels of critical cytokines (IL-2 and IL-18) leading to CMI loss regulation.

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