Development of cathepsin-L cysteine proteinase based Dot-enzyme-linked immunosorbent assay for the diagnosis of Fasciola gigantica infection in buffaloes

Varghese, Anju; Raina, O.K.; Nagar, Gaurav; Garg, Rajat; Banerjee, P. S.; Maharana, Biswa Ranjan; Kollannur, Justin D.

doi:10.1016/j.vetpar.2011.07.032

Cited by 21 publications

(22 citation statements)

References 21 publications

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“…Therefore, more sensitive diagnostic methods are required. The dot‐enzyme‐linked immunosorbent assay (dot‐ELISA) is a simple, rapid and scalable process to detect antigens or antibodies while screening large numbers of samples at once . Serological diagnosis including traditional ELISA and dot‐ELISA has been used to detect psoroptic sheep scab around the world .…”

Section: Introductionmentioning

confidence: 99%

Evaluating troponin C from Psoroptes cuniculi as a diagnostic antigen for a dot‐ELISA assay to diagnose mite infestations in rabbits

Zheng

Zhang

et al. 2014

Parasite Immunology

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The mite Psoroptes cuniculi is globally widespread and has a serious impact on commercial rabbit breeding. In China, diagnosis of P. cuniculi is currently based on conventional clinical methods that entail numerous disadvantages, including their failure to diagnose subclinical infections. Hence, alternative measures are required, and dot-ELISA is one of the most promising strategies. We cloned and expressed the recombinant P. cuniculi troponin C gene for use as a basis for novel dot-ELISA assay to detect P. cuniculi infections in rabbits. This amplified sequence encoded a 153 amino acid protein of 17·6 kDa and theoretical pI 4·18 without signal peptide. The recombinant troponin C of P. cuniculi is an outer membrane protein and may also be a new P. cuniculi allergen. Results of dot-ELISA test showed that this novel assay had more than 90% sensitivity but low specificity in distinguishing infections with P. cuniculi or Sarcoptic scabiei, despite very high agreement between observers (97-99%; κ values ranged from 0·95 to 0·98 for inter- and intra-observer variability test). This study showed that this novel method, at present, lacks diagnostic utility. Therefore, although simple serological assays such as dot-ELISA show great promise as diagnostic tools, we suggest that troponin C is not a suitable diagnostic antigen candidate.

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Section: Introductionmentioning

confidence: 99%

Evaluating troponin C from Psoroptes cuniculi as a diagnostic antigen for a dot‐ELISA assay to diagnose mite infestations in rabbits

Zheng

Zhang

et al. 2014

Parasite Immunology

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“…Recently, native cathepsin-L cysteine proteinase was purified from the excretory secretory products of Fasciola and used for sero-diagnosis of Fasciola infection in buffaloes by Dot -ELISA. The results demonstrated that cathepsin-L cysteine proteinase based Dot-ELISA achieved 90.00 % sensitivity and 100 % specificity (Varghese et al 2012). Our data indicated when ES antigens of F. gigantica used for i-ELISA or Dot-ELISA designing, cross-reaction was not detected.…”

Section: Resultsmentioning

confidence: 74%

Comparison between in-house indirect ELISA and Dot-ELISA for the diagnosis of Fasciola gigantica in cattle

et al. 2015

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This survey was done to investigate the efficacy of the in-house indirect ELISA (iELISA) and Dot-ELISA methods Prepared from excretion-secretory (ES Ag) and Crude (Cr Ag) antigens of Fasciola for sero-diagnosis of Fasciola gigantica in cattle. The liver specimens of slaughtered cattle were collected and their liver examined macroscopically and microscopically for infestation to fasciolosis. Sera from two groups of cattle, one infected with fasciolosis (n = 60) and the other non-infected with fasciolosis (n = 60), were used in the iELISA and Dot-ELISA test; grouping based on histopathology results. Except specificity, other parameters such as, sensitivity, accuracy, positive and negative predictive values of both Dot-ELISA and iELISA done with ES Ag were better than those of tests performed with Cr Ag. Interestingly, the reliability of two methods was very good similar to one another.

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“…Test sensitivity was calculated as: sensitivity = ELISA positive/true positive × 100%. Test specificity was calculated as: specificity = ELISA negative/true negative × 100% [31]. …”

Section: Methodsmentioning

confidence: 99%

Characterisation and analysis of thioredoxin peroxidase as a potential antigen for the serodiagnosis of sarcoptic mange in rabbits by dot-ELISA

Zhang

Zheng

et al. 2013

BMC Infect Dis

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BackgroundScabies caused by Sarcoptes scabiei is a widespread but a neglected tropical zoonosis. In this study, we characterised a S. scabiei thioredoxin peroxidase (SsTPx) and evaluated a recombinant SsTPx as a diagnostic antigen in rabbits.MethodsThe open reading frame of the gene encoding SsTPx-2 was amplified and the recombinant protein was expressed in Escherichia coli cells and purified. SsTPx was localized in mite tissue by immunolocalisation using the purified recombinant protein. Serodiagnosis assays were carried out in 203 New Zealand White rabbit serum samples by dot-ELISA.ResultThe open reading frame (489 bp) of the gene encodes an 18.11 kDa protein, which showed highly homology to that of Psoroptes cuniculi (98.77% identity) and belongs to the 2-Cys family of peroxiredoxins. SsTPx was mainly distributed in muscle tissues of mites, integument of the epidermis and the anterior end of S. scabiei. Although SsTPx cross-reactivity with psoroptic mites was observed, the SsTPx dot-ELISA showed excellent diagnostic ability, with 95.3% sensitivity and 93.8% specificity in mange-infected and uninfected groups.ConclusionsThis study showed that the purified SsTPx is a highly sensitive antigen for the diagnosis of mange infection by dot-ELISA. This technique is a rapid and convenient method that can be used worldwide for the clinical diagnosis of sarcoptic mange in rabbits, and is especially useful in developing regions.

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Development of cathepsin-L cysteine proteinase based Dot-enzyme-linked immunosorbent assay for the diagnosis of Fasciola gigantica infection in buffaloes

Cited by 21 publications

References 21 publications

Evaluating troponin C from Psoroptes cuniculi as a diagnostic antigen for a dot‐ELISA assay to diagnose mite infestations in rabbits

Evaluating troponin C from Psoroptes cuniculi as a diagnostic antigen for a dot‐ELISA assay to diagnose mite infestations in rabbits

Comparison between in-house indirect ELISA and Dot-ELISA for the diagnosis of Fasciola gigantica in cattle

Characterisation and analysis of thioredoxin peroxidase as a potential antigen for the serodiagnosis of sarcoptic mange in rabbits by dot-ELISA

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