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

Zhang R, Zheng W, Wu X, Jise Q, Ren Y, Nong X, Gu X, Wang S, Peng X, Lai S, Yang G - BMC Infect. Dis. (2013)

Bottom Line: 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.This study showed that the purified SsTPx is a highly sensitive antigen for the diagnosis of mange infection by dot-ELISA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.

ABSTRACT

Background: Scabies 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.

Methods: The 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.

Result: The 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.

Conclusions: This 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.

Show MeSH

Related in: MedlinePlus

Dot-ELISA showing reactivity of infected group and uninfected groups using three concentrations of recombinant SsTPx protein as a capture antigen. Lane 1, 2 μg/mL protein; Lane 2, 6 μg/mL protein; Lane 3, 24 μg/mL protein.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3733899&req=5

Figure 4: Dot-ELISA showing reactivity of infected group and uninfected groups using three concentrations of recombinant SsTPx protein as a capture antigen. Lane 1, 2 μg/mL protein; Lane 2, 6 μg/mL protein; Lane 3, 24 μg/mL protein.

Mentions: The optimal dilution of the serum and controls for use in dot-ELISA assays was 1:200 and the optimal concentration of the recombinant SsTPx antigen was 2 μg/mL (data not shown). A total of 203 rabbit sera were evaluated. Most experimentally or naturally infected S. scabiei sera (average 81/85, 95.3% sensitivity) showed positive reactivity in dot-ELISAs, while most S. scabiei-negative sera (average 111/118, 93.8% specificity) showed no reaction with SsTPx. In the control groups, SsTPx showed no cross-reaction with E. stiedae (0/6) and T. pisiformis (0/6). Cross-reactivity was observed with P. cuniculi-infected (7–8/12) serum although the definition of the dots was less clear that those of the S. scabiei infected group (Figure 4). No obvious variation was observed among the results for the three time-points tested indicating the stability of the membrane-bound antigen.


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

Zhang R, Zheng W, Wu X, Jise Q, Ren Y, Nong X, Gu X, Wang S, Peng X, Lai S, Yang G - BMC Infect. Dis. (2013)

Dot-ELISA showing reactivity of infected group and uninfected groups using three concentrations of recombinant SsTPx protein as a capture antigen. Lane 1, 2 μg/mL protein; Lane 2, 6 μg/mL protein; Lane 3, 24 μg/mL protein.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3733899&req=5

Figure 4: Dot-ELISA showing reactivity of infected group and uninfected groups using three concentrations of recombinant SsTPx protein as a capture antigen. Lane 1, 2 μg/mL protein; Lane 2, 6 μg/mL protein; Lane 3, 24 μg/mL protein.
Mentions: The optimal dilution of the serum and controls for use in dot-ELISA assays was 1:200 and the optimal concentration of the recombinant SsTPx antigen was 2 μg/mL (data not shown). A total of 203 rabbit sera were evaluated. Most experimentally or naturally infected S. scabiei sera (average 81/85, 95.3% sensitivity) showed positive reactivity in dot-ELISAs, while most S. scabiei-negative sera (average 111/118, 93.8% specificity) showed no reaction with SsTPx. In the control groups, SsTPx showed no cross-reaction with E. stiedae (0/6) and T. pisiformis (0/6). Cross-reactivity was observed with P. cuniculi-infected (7–8/12) serum although the definition of the dots was less clear that those of the S. scabiei infected group (Figure 4). No obvious variation was observed among the results for the three time-points tested indicating the stability of the membrane-bound antigen.

Bottom Line: 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.This study showed that the purified SsTPx is a highly sensitive antigen for the diagnosis of mange infection by dot-ELISA.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Ya'an 625014, China.

ABSTRACT

Background: Scabies 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.

Methods: The 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.

Result: The 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.

Conclusions: This 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.

Show MeSH
Related in: MedlinePlus