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Identification of immunodominant antigens in canine leptospirosis by Multi-Antigen Print ImmunoAssay (MAPIA).

Thomé S, Lessa-Aquino C, Ko AI, Lilenbaum W, Medeiros MA - BMC Vet. Res. (2014)

Bottom Line: The data were analyzed and ROC curves were generated.Altogether, LigB [131-649aa] L. interrogans Canicola, LigB [131-649aa] L. kirschneri Gryppotyphosa and LipL32 L. interrogans Copenhageni showed best accuracy (AUC = 0.826 to 0.869), with 70% specificity and sensitivity ranging from 89% to 95%.These results reinforce their potential as diagnostic candidates for the development of new methods for the serological diagnosis of canine leptospirosis.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Bacteriology, Department of Microbiology and Parasitology, Universidade Federal Fluminense, Niterói, RJ, Brazil. sabrina_thome@vm.uff.br.

ABSTRACT

Background: The microscopic agglutination test (MAT), the standard method for serological diagnosis of leptospirosis, may present limitations regarding its sensitivity. Current studies suggest that Leptospira immunoglobulin-like (Lig) proteins and LipL32 are of particular interest as serodiagnostic markers since they are present only in pathogenic species of the Leptospira genus. The purpose of this study was to identify leptospiral immunodominant proteins that are recognized by canine sera from diseased dogs.

Results: A total of 109 dogs were studied, including seroreactive dogs (MAT ≥800) and dogs with no seroreactivity detectable by MAT. Eight recombinant fragments (31-70 kDa) of pathogenic Leptospira were tested for their use as diagnostic markers for canine leptospirosis using the Multi-antigen Print Immunoassay (MAPIA) platform: LigB [582-947aa] from L. interrogans serovar Pomona, L. interrogans serovar Copenhageni and L. kirschneri serovar Gryppotyphosa, LigB [131-649aa] from L. interrogans serovar Copenhageni, L. interrogans serovar Canicola and L. kirschneri serovar Gryppotyphosa, LigA [625-1224aa] L. interrogans serovar Copenhageni and LipL32 from L. interrogans serovar Copenhageni. The data were analyzed and ROC curves were generated. Altogether, LigB [131-649aa] L. interrogans Canicola, LigB [131-649aa] L. kirschneri Gryppotyphosa and LipL32 L. interrogans Copenhageni showed best accuracy (AUC = 0.826 to 0.869), with 70% specificity and sensitivity ranging from 89% to 95%.

Conclusions: These results reinforce their potential as diagnostic candidates for the development of new methods for the serological diagnosis of canine leptospirosis.

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Related in: MedlinePlus

Box plots representing the overall reactivity detected for each group of samples against the recombinant proteins. Bars indicate the maximum or minimum values; the purple area indicates the 25th percentile; the green area indicates de 75th percentile; the line between purple and green areas indicates de median. Sample groups: P1, seropositive dogs; N2, housed vaccinated dogs; N3, stray dogs; N4, dogs with other febrile syndromes.
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Fig2: Box plots representing the overall reactivity detected for each group of samples against the recombinant proteins. Bars indicate the maximum or minimum values; the purple area indicates the 25th percentile; the green area indicates de 75th percentile; the line between purple and green areas indicates de median. Sample groups: P1, seropositive dogs; N2, housed vaccinated dogs; N3, stray dogs; N4, dogs with other febrile syndromes.

Mentions: The purified recombinant proteins used for printing the MAPIA strips were analyzed by SDS PAGE 12.5% and showed high homogeneity (Figure 1). The MAPIA densitometry results showed that, in general, group N2 (housed vaccinated dogs) had lower IgM background against the recombinant proteins than groups N3 (stray dogs) and N4 (dogs with other febrile syndromes) and group P1 (seropositive dogs) had a stronger IgM reactivity (Figure 2). The reactivity detected for group P1 (seropositive dogs) against LigB [131-649aa] L. kirschneri Gryppotyphosa, LigB [131-649aa] L. interrogans Canicola and LipL32 L. interrogans Copenhageni were the most intense, with a higher median reactivity when compared to the control groups (Figure 2).Figure 1


Identification of immunodominant antigens in canine leptospirosis by Multi-Antigen Print ImmunoAssay (MAPIA).

Thomé S, Lessa-Aquino C, Ko AI, Lilenbaum W, Medeiros MA - BMC Vet. Res. (2014)

Box plots representing the overall reactivity detected for each group of samples against the recombinant proteins. Bars indicate the maximum or minimum values; the purple area indicates the 25th percentile; the green area indicates de 75th percentile; the line between purple and green areas indicates de median. Sample groups: P1, seropositive dogs; N2, housed vaccinated dogs; N3, stray dogs; N4, dogs with other febrile syndromes.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4269070&req=5

Fig2: Box plots representing the overall reactivity detected for each group of samples against the recombinant proteins. Bars indicate the maximum or minimum values; the purple area indicates the 25th percentile; the green area indicates de 75th percentile; the line between purple and green areas indicates de median. Sample groups: P1, seropositive dogs; N2, housed vaccinated dogs; N3, stray dogs; N4, dogs with other febrile syndromes.
Mentions: The purified recombinant proteins used for printing the MAPIA strips were analyzed by SDS PAGE 12.5% and showed high homogeneity (Figure 1). The MAPIA densitometry results showed that, in general, group N2 (housed vaccinated dogs) had lower IgM background against the recombinant proteins than groups N3 (stray dogs) and N4 (dogs with other febrile syndromes) and group P1 (seropositive dogs) had a stronger IgM reactivity (Figure 2). The reactivity detected for group P1 (seropositive dogs) against LigB [131-649aa] L. kirschneri Gryppotyphosa, LigB [131-649aa] L. interrogans Canicola and LipL32 L. interrogans Copenhageni were the most intense, with a higher median reactivity when compared to the control groups (Figure 2).Figure 1

Bottom Line: The data were analyzed and ROC curves were generated.Altogether, LigB [131-649aa] L. interrogans Canicola, LigB [131-649aa] L. kirschneri Gryppotyphosa and LipL32 L. interrogans Copenhageni showed best accuracy (AUC = 0.826 to 0.869), with 70% specificity and sensitivity ranging from 89% to 95%.These results reinforce their potential as diagnostic candidates for the development of new methods for the serological diagnosis of canine leptospirosis.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Bacteriology, Department of Microbiology and Parasitology, Universidade Federal Fluminense, Niterói, RJ, Brazil. sabrina_thome@vm.uff.br.

ABSTRACT

Background: The microscopic agglutination test (MAT), the standard method for serological diagnosis of leptospirosis, may present limitations regarding its sensitivity. Current studies suggest that Leptospira immunoglobulin-like (Lig) proteins and LipL32 are of particular interest as serodiagnostic markers since they are present only in pathogenic species of the Leptospira genus. The purpose of this study was to identify leptospiral immunodominant proteins that are recognized by canine sera from diseased dogs.

Results: A total of 109 dogs were studied, including seroreactive dogs (MAT ≥800) and dogs with no seroreactivity detectable by MAT. Eight recombinant fragments (31-70 kDa) of pathogenic Leptospira were tested for their use as diagnostic markers for canine leptospirosis using the Multi-antigen Print Immunoassay (MAPIA) platform: LigB [582-947aa] from L. interrogans serovar Pomona, L. interrogans serovar Copenhageni and L. kirschneri serovar Gryppotyphosa, LigB [131-649aa] from L. interrogans serovar Copenhageni, L. interrogans serovar Canicola and L. kirschneri serovar Gryppotyphosa, LigA [625-1224aa] L. interrogans serovar Copenhageni and LipL32 from L. interrogans serovar Copenhageni. The data were analyzed and ROC curves were generated. Altogether, LigB [131-649aa] L. interrogans Canicola, LigB [131-649aa] L. kirschneri Gryppotyphosa and LipL32 L. interrogans Copenhageni showed best accuracy (AUC = 0.826 to 0.869), with 70% specificity and sensitivity ranging from 89% to 95%.

Conclusions: These results reinforce their potential as diagnostic candidates for the development of new methods for the serological diagnosis of canine leptospirosis.

Show MeSH
Related in: MedlinePlus