scholarly journals Identification and Diagnostic Utility of Leishmania infantum Proteins Found in Urine Samples from Patients with Visceral Leishmaniasis

2012 ◽  
Vol 19 (6) ◽  
pp. 935-943 ◽  
Author(s):  
Claudia Abeijon ◽  
Suely S. Kashino ◽  
Fernando O. Silva ◽  
Dorcas L. Costa ◽  
Ricardo T. Fujiwara ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Infantum ◽  
Antigen Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Accession Number ◽  
Content Type ◽  
Link Type ◽  
Detection Assay ◽  
Ncbi Accession ◽  
Ncbi Accession Number

ABSTRACTDespite the clear need to control visceral leishmaniasis (VL), the existing diagnostic tests have serious shortcomings. Here, we introduce an innovative approach to directly identifyLeishmania infantumantigens producedin vivoin humans with VL. We combined reverse-phase high-performance liquid chromatography (RP-HPLC) with mass spectrometry and categorized three distinctL. infantumproteins presumably produced in bone marrow/spleen/liver and excreted in the urine of patients with VL. The genes coding for these proteins (L. infantumiron superoxide dismutase, NCBI accession numberXP_001467866.1;L. infantumtryparedoxin, NCBI accession numberXP_001466642.1; andL. infantumnuclear transport factor 2, NCBI accession numberXP_001463738.1) were cloned, and the recombinant molecules were produced inEscherichia coli. Antibodies to these proteins were produced in rabbits and chickens and were used to develop a capture enzyme-linked immunosorbent assay (ELISA) designed to detect theseL. infantumantigens in the urine of VL patients. Specificity of the antibodies was confirmed by a Western blot analysis using both recombinant proteins and whole parasite extract. Importantly, a urinary antigen detection assay assembled with pairs of antibodies specific for each of these antigens identified 17 of 19 patients with VL. These results indicate that an improved antigen detection assay based onL. infantumproteins present in the urine of patients with VL may represent an important new strategy for the development of a specific and accurate diagnostic test that has the potential to both distinguish active VL from asymptomatic infection and serve as an important tool to monitor therapy efficacy.

scholarly journals Subtractive Phage Display Selection from Canine Visceral Leishmaniasis Identifies Novel Epitopes That Mimic Leishmania infantum Antigens with Potential Serodiagnosis Applications

2013 ◽  
Vol 21 (1) ◽  
pp. 96-106 ◽  
Author(s):  
Lourena E. Costa ◽  
Mayara I. S. Lima ◽  
Miguel A. Chávez-Fumagalli ◽  
Daniel Menezes-Souza ◽  
Vivian T. Martins ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Phage Display ◽  
Leishmania Infantum ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Phage Display Technology ◽  
Predictive Values ◽  
Content Type ◽  
Phage Elisa

ABSTRACTVisceral leishmaniasis (VL) is a zoonotic disease that is endemic to Brazil, where dogs are the main domestic parasite reservoirs, and the percentages of infected dogs living in regions where canine VL (CVL) is endemic have ranged from 10% to 62%. Despite technological advances, some problems have been reported with CVL serodiagnosis. The present study describes a sequential subtractive selection through phage display technology from polyclonal antibodies of negative and positive sera that resulted in the identification of potential bacteriophage-fused peptides that were highly sensitive and specific to antibodies of CVL. A negative selection was performed in which phage clones were adhered to purified IgGs from healthy andTrypanosoma cruzi-infected dogs to eliminate cross-reactive phages. The remaining supernatant nonadhered phages were submitted to positive selection against IgG from the blood serum of dogs that were infected withLeishmania infantum. Phage clones that adhered to purified IgGs from the CVL-infected serum samples were selected. Eighteen clones were identified and their reactivities tested by a phage enzyme-linked immunosorbent assay (phage-ELISA) against the serum samples from infected dogs (n= 31) compared to those from vaccinated dogs (n= 21), experimentally infected dogs with cross-reactive parasites (n= 23), and healthy controls (n= 17). Eight clones presented sensitivity, specificity, and positive and negative predictive values of 100%, and they showed no cross-reactivity withT. cruzi- orEhrlichia canis-infected dogs or with dogs vaccinated with two different commercial CVL vaccines in Brazil. Our study identified eight mimotopes ofL. infantumantigens with 100% accuracy for CVL serodiagnosis. The use of these mimotopes by phage-ELISA proved to be an excellent assay that was reproducible, simple, fast, and inexpensive, and it can be applied in CVL-monitoring programs.

scholarly journals Genome Sequence of “Candidatus Rickettsia colombianensi,” a Novel Tick-Associated Bacterium Distributed in Colombia

2019 ◽  
Vol 8 (14) ◽  
Author(s):  
Jorge Miranda ◽  
Salim Mattar ◽  
Andrés Puerta-González ◽  
Carlos Muskus ◽  
José A. Oteo
Keyword(s):  
Genome Sequence ◽  
Draft Genome ◽  
Accession Number ◽  
Content Type ◽  
First Report ◽  
Rickettsia Monacensis ◽  
Ncbi Accession ◽  
Rickettsia Heilongjiangensis ◽  
Ncbi Accession Number

This is the first report of the genome sequence of “Candidatus Rickettsia colombianensi” strain Adcor 2, deposited in DDBJ/ENA/GenBank under the accession number RAQN00000000. The draft genome showed 36.01% similarity with that of Rickettsia monacensis strain IrR/Munich (NCBI accession number LN794217), 37.81% similarity with that of Rickettsia heilongjiangensis 054 (NCBI accession number CP002912), and 43.88% similarity with that of Rickettsia tamurae AT-1 (NCBI accession number CCMG01000001).

scholarly journals Phylo-geo-network and haplogroup analysis of 611 novel coronavirus (SARS-CoV-2) genomes from India

2021 ◽  
Vol 4 (5) ◽  
pp. e202000925
Author(s):  
Rezwanuzzaman Laskar ◽  
Safdar Ali
Keyword(s):  
Viral Evolution ◽  
Reference Sequence ◽  
The Novel ◽  
Accession Number ◽  
Global Epidemic ◽  
The Core ◽  
Link Type ◽  
Ncbi Accession ◽  
Novel Coronavirus ◽  
Ncbi Accession Number

The novel coronavirus (SARS-CoV-2) from Wuhan China discovered in December 2019 has since developed into a global epidemic. Presently, we constructed and analyzed the phylo-geo-network of SARS-CoV-2 genomes from across India to understand the viral evolution in the country. A total of 611 full-length genomes from different states of India were extracted from the EpiCov repository of GISAID initiative on 6 June, 2020. Their alignment with the reference sequence (Wuhan, NCBI accession number NC_045512.2) uncovered 270 parsimony informative sites. Furthermore, 339 genomes were divided into 51 haplogroups. The network revealed the core haplogroup as that of reference sequence NC_045512.2 (Haplogroup A1) with 157 identical sequences present across 16 states. Remaining haplogroups had <10 identical sequences across a maximum of three states. Some states with fewer samples had more haplogroups. Forty-one haplogroups were localized exclusively to any one state. The two most common lineages are B6 and B1 (Pangolin) whereas clade A2a (Covidex) appears to be the most predominant in India. Because the pandemic is still emerging, the observations need to be monitored.

scholarly journals Development of a Multiplexed Assay for Detection ofLeishmania donovaniandLeishmania infantumProtein Biomarkers in Urine Samples of Patients with Visceral Leishmaniasis

2019 ◽  
Vol 57 (5) ◽  
Author(s):  
Claudia Abeijon ◽  
Fabiana Alves ◽  
Severine Monnerat ◽  
Monique Wasunna ◽  
Jane Mbui ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Diagnostic Test ◽  
Leishmania Donovani ◽  
Protein Biomarkers ◽  
Serological Tests ◽  
Assay Sensitivity ◽  
Content Type ◽  
Detection Assay ◽  
Previously Treated ◽  
New Biomarkers

ABSTRACTVisceral leishmaniasis (VL) is a serious and fatal disease caused by the parasitesLeishmania infantumandLeishmania donovani. The gold standard diagnostic test for VL is the demonstration of parasites or their DNA in spleen, lymph node, or bone marrow aspirates. Serological tests exist but cannot distinguish active VL from either prior exposure to the parasites or previously treated VL disease. Using mass spectroscopy, we have previously identified threeL. infantumprotein biomarkers (Li-isd1,Li-txn1, andLi-ntf2) in the urine of VL patients and developed a sensitive and specific urine-based antigen detection assay for the diagnosis of VL that occurs in Brazil (where VL is caused byL. infantum). However, unpublished observations from our laboratory at DetectoGen showed that these biomarkers were detected in only 55% to 60% of VL patients from India and Kenya, where the disease is caused byL. donovani. Here, we report the discovery and characterization of two new biomarkers ofL. donovani(Ld-mao1andLd-ppi1) present in the urine of VL patients from these two countries. Capture enzyme-linked immunosorbent assays using specific rabbit IgG and chicken IgY were developed, and the assays had sensitivities of 44.4% and 28.8% for the detection ofLd-mao1andLd-ppi1, respectively. In contrast, a multiplexed assay designed to simultaneously detect all five leishmanial biomarkers markedly increased the assay sensitivity to 82.2%. These results validate the utility of leishmanial protein biomarkers found in the urine of VL patients as powerful tools for the development of an accurate diagnostic test for this disease.

scholarly journals Immune Responses of Iranian Patients with Visceral Leishmaniasis and Recovered Individuals to LCR1 of Leishmania infantum

2014 ◽  
Vol 21 (4) ◽  
pp. 518-525 ◽  
Author(s):  
Hamid M. Niknam ◽  
Firoozeh Abrishami ◽  
Mohammad Doroudian ◽  
Mosayeb Rostamian ◽  
Maryam Moradi ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Immune Responses ◽  
Leishmania Infantum ◽  
Mononuclear Cells ◽  
Antibody Responses ◽  
Antibody Detection ◽  
Recent History ◽  
Content Type ◽  
History Of ◽  
Ifn Γ

ABSTRACTVisceral leishmaniasis is a serious public health problem.Leishmania infantumis one of its causative agents. LCR1 is an immunogen fromL. infantum. Antibodies against this protein have been detected in visceral leishmaniasis patients. The aim of this study was to define the antibody and cellular immune responses against LCR1 in Iranian visceral leishmaniasis patients and recovered individuals. The LCR1 protein was produced in recombinant form. Antibody responses against this protein were studied in Iranian individuals with a recent history of visceral leishmaniasis. Responses of peripheral blood mononuclear cells to this protein were studied in Iranian individuals who had recovered from visceral leishmaniasis. Our data show that (i) there was an antibody response to LCR1 in each individual with a recent history of visceral leishmaniasis studied, (ii) there was neither a proliferative response nor production of gamma interferon (IFN-γ) or interleukin 10 in response to LCR1 by mononuclear cells from individuals who had recovered from visceral leishmaniasis, and (iii) individuals who have recovered from visceral leishmaniasis show ongoing immune responses long after recovery from the disease. These data show that there are no detectable cellular memory responses to LCR1 in Iranian individuals who have recovered from visceral leishmaniasis, while there are detectable antibody responses in patients with this disease. Our data suggest that LCR1 has potential applications for the diagnosis of leishmaniasis through antibody detection, while the application of LCR1 alone for induction of IFN-γ in individuals who recovered from this disease is not supported. The presence of long-lasting immune reactivities in individuals who recovered from the disease may show the necessity of extended medical surveillance for these individuals.

scholarly journals Immunodetection of Fasciola gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis

2013 ◽  
Vol 20 (10) ◽  
pp. 1569-1577 ◽  
Author(s):  
Abdelfattah M. Attallah ◽  
Faisal A. Bughdadi ◽  
Atef M. El-Shazly ◽  
Hisham Ismail
Keyword(s):  
Laboratory Diagnosis ◽  
Fasciola Gigantica ◽  
Antigen Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Target Antigen ◽  
Immunoperoxidase Staining ◽  
Serum Samples ◽  
Content Type ◽  
Human Fascioliasis ◽  
Circulating Antigen

ABSTRACTCurrently, the laboratory diagnosis of human fascioliasis is based on the parasitological examination of parasite eggs in stool specimens and serological detection of specific antibodies in serum samples, which are often unreliable diagnostic approaches. Ideally, a sensitive and specific diagnostic test forFasciolainfection should be based on the detection of circulatingFasciolaantigen, which implies active infection. Here, a 27-kDa-molecular-mass antigen was identified in aFasciola giganticaadult worm antigen preparation, excretory-secretory products, and sera fromF. gigantica-infected individuals, and it was not detected in antigenic extracts of other parasites and sera from noninfected individuals. The target antigen was isolated and partially characterized as a protein. Immunoperoxidase staining located the target epitope within teguments and guts ofF. giganticaadult worms. The performance characteristics of a newly developed enzyme-linked immunosorbent assay (ELISA) based onF. giganticacirculating antigen detection in serum (FgCA-27 ELISA) were investigated using sera of 120 parasitologically diagnosedF. gigantica-infected individuals and 80 noninfected individuals. The area under the receiving operating characteristic (ROC) curve (AUC) for ELISA was significantly high (AUC = 0.961,P< 0.0001) for discriminatingFasciola-infected and noninfected individuals. The developed assay showed high degrees of sensitivity, specificity, and efficiency (>93%), and a significant correlation (r= 0.715,P< 0.0001) between antigen level and parasite egg count was shown. In conclusion, a 27-kDaFasciolaantigen was identified in sera ofF. gigantica-infected individuals. A highly sensitive and specificFasciolaantigen detection assay, FgCA-27 ELISA, was developed for laboratory diagnosis of human fascioliasis.

scholarly journals Identification of Mycobacterium tuberculosis Ornithine Carboamyltransferase in Urine as a Possible Molecular Marker of Active Pulmonary Tuberculosis

2008 ◽  
Vol 15 (4) ◽  
pp. 638-643 ◽  
Author(s):  
Danielle R. Napolitano ◽  
Nira Pollock ◽  
Suely S. Kashino ◽  
Virmondes Rodrigues ◽  
Antonio Campos-Neto
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Pulmonary Tuberculosis ◽  
Mononuclear Cells ◽  
Antigen Detection ◽  
Active Tuberculosis ◽  
Urine Samples ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Peripheral Blood Mononuclear ◽  
Detection Assay

ABSTRACT Although the antigen detection assay has the potential to discriminate active tuberculosis from latent infection, development of such a test for the accurate diagnosis of this serious disease has only recently become a matter of interest. Here we present evidence that a Mycobacterium tuberculosis protein (ornithine carboamyltransferase, coded for by MT_1694; Rv1656 [argF]) is an interesting candidate molecule for this test development. The protein was initially discovered by mass spectroscopy in urine of patients with pulmonary tuberculosis and shown by Western blot analysis to be present in M. tuberculosis crude cell extract as well as in the culture supernatant (“secreted” protein). In addition, a recombinant ornithine carboamyltransferase (rMT1694) produced in Escherichia coli was recognized by immunoglobulin G (IgG) antibodies from patients with active tuberculosis but not by IgG from uninfected healthy subjects. Moreover, rMT1694 was strongly recognized by peripheral blood mononuclear cells from both healthy tuberculin purified protein derivative (PPD)-positive individuals and patients with pulmonary tuberculosis. More importantly, a capture enzyme-linked immunosorbent assay formatted with rabbit IgG antibodies specific to rMT1694 was able to identify the presence of this antigen in urine samples from 6 of 16 patients with pulmonary tuberculosis and in none of 16 urine samples collected from healthy PPD+ controls. These results indicate that an improved antigen detection assay based on M. tuberculosis ornithine carboamyltransferase may represent an important new strategy for the development of a specific and accurate diagnostic test for tuberculosis.

scholarly journals Whole-Genome Sequence of an Indian Group A Streptococcus emm Type 1-2 Strain Isolated from a Blood Sample in North India

2020 ◽  
Vol 9 (19) ◽  
Author(s):  
Vivek Sagar ◽  
Anuradha Chakraborti ◽  
Rajesh Kumar
Keyword(s):  
Partial Information ◽  
Group A Streptococcus ◽  
North India ◽  
Whole Genome Sequence ◽  
Accession Number ◽  
Genetic Characteristics ◽  
Ncbi Accession ◽  
Group A ◽  
Ncbi Accession Number

Group A Streptococcus emm type 1-2 is more prevalent than emm type 1 in India. Only partial information is available about the genetic characteristics of this type. Here, genome sequencing of emm type 1-2 strain 1085 (from blood) was conducted. A contig 2,010,300 bp long, with a total of 1,877 annotated proteins, was obtained (NCBI accession number CP047120, assembly accession number ASM983284v1).

scholarly journals Shewanella algae and Microbulbifer elongatus from marine macro-algae – isolation and characterization of agar-hydrolysing bacteria

2020 ◽  
Vol 2 (11) ◽  
Author(s):  
Keerthana Ponni Kandasamy ◽  
Radhesh Krishnan Subramanian ◽  
Radhakrishnan Srinivasan ◽  
Sengali Ragunath ◽  
G. Balaji ◽  
...  
Keyword(s):  
Type Species ◽  
Degradation Products ◽  
Accession Number ◽  
Content Type ◽  
Link Type ◽  
Isolation And Characterization ◽  
Shewanella Algae ◽  
Agarase Activity ◽  
Macro Algae

Macro-algae are a good source of agar oligosaccharides, which can be obtained through bacterial enzymatic hydrolysis. The agarase enzyme secreted by the micro-organisms cleaves the cell wall of the algae and releases agar oligosaccharides as degradation products with various applications. Agarolytic bacteria were isolated from the marine algae Kappaphycus sp., and Sargassum sp., and studied for their agar-degrading properties. Among the 70 isolates, 2 isolates (A13 and Sg8) showed agarase activity in in vitro assays. The maximum agarolytic index was recorded in the isolate Sg8 (3.75 mm and 4.29 µg ml−1 agarase activity), followed by the isolate A13 (2.53 mm and 2.6 µg ml−1 agarase activity). Optimum agarase production of isolate Sg8 was observed at pH7 and at a temperature of 25 °C in 24–48 h, whereas for isolate A13 the optimum production was at pH7 and at a temperature of 37 °C in 48 h. The identities of the agarolytic isolates (Sg8 and A13) were confirmed based on microscopy, morphological, biochemical and molecular analysis as Shewanella algae [National Center for Biotechnology Information (NCBI) GenBank accession number MK121204.1] and Microbulbifer elongatus [NCBI GenBank accession number MK825484.1], respectively.

Sign in / Sign up

Export Citation Format

Share Document