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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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ROC curves showing the diagnostic accuracy of each recombinant protein analyzed individually (A) or in combination (B) when combining the 3 control groups N2, N3 and N4 as a single group. In A, proteins are sorted from left to right by decreasing AUC values. The combination of the first 3 antigens (B, dark brown) provides best accuracy. Sample groups: P1, seropositive dogs; N2, housed vaccinated dogs; N3, stray dogs; N4, dogs with other febrile syndromes.
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Fig3: ROC curves showing the diagnostic accuracy of each recombinant protein analyzed individually (A) or in combination (B) when combining the 3 control groups N2, N3 and N4 as a single group. In A, proteins are sorted from left to right by decreasing AUC values. The combination of the first 3 antigens (B, dark brown) provides best accuracy. Sample groups: P1, seropositive dogs; N2, housed vaccinated dogs; N3, stray dogs; N4, dogs with other febrile syndromes.

Mentions: In order to assess the diagnostic accuracy of the recombinant proteins, individual antigen ROC curves were generated and the corresponding AUC was determined. Antigens were then ranked by decreasing AUC and multiple antigens ROC curves were generated. Figure 3A shows the individual ROC curves for each of the recombinant proteins when we compared the seropositive dogs (group P1) against all the other groups combined as a single group (groups N2, N3 and N4). The AUC values are shown in Table 1. LipL32 L. interrogans Copenhageni provided best accuracy (AUC = 0.869), followed by LigB [131-649aa] L. kirschneri Gryppotyphosa (AUC = 0.855) and LigB [131-649aa] L. interrogans Canicola (AUC = 0.826). Furthermore, when group P1 was compared to groups N2, N3 and N4 separately, the same antigens showed best performance. The combination of those three antigens improved the diagnostic accuracy when considering all control samples in a single group (Figure 3B) and also when group P1 was compared to group N3 (stray dogs, Figure 4). Using group N2 as the control group, LipL32 and LigB [131-649aa] L. kirschneri Gryppotyphosa provided the best performance. When we consider group N4 (dogs with other febrile syndromes) as control, however, the addition of extra antigens did not impact the diagnostic performance of LipL32 (Figure 4).Figure 3


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)

ROC curves showing the diagnostic accuracy of each recombinant protein analyzed individually (A) or in combination (B) when combining the 3 control groups N2, N3 and N4 as a single group. In A, proteins are sorted from left to right by decreasing AUC values. The combination of the first 3 antigens (B, dark brown) provides best accuracy. 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

Fig3: ROC curves showing the diagnostic accuracy of each recombinant protein analyzed individually (A) or in combination (B) when combining the 3 control groups N2, N3 and N4 as a single group. In A, proteins are sorted from left to right by decreasing AUC values. The combination of the first 3 antigens (B, dark brown) provides best accuracy. Sample groups: P1, seropositive dogs; N2, housed vaccinated dogs; N3, stray dogs; N4, dogs with other febrile syndromes.
Mentions: In order to assess the diagnostic accuracy of the recombinant proteins, individual antigen ROC curves were generated and the corresponding AUC was determined. Antigens were then ranked by decreasing AUC and multiple antigens ROC curves were generated. Figure 3A shows the individual ROC curves for each of the recombinant proteins when we compared the seropositive dogs (group P1) against all the other groups combined as a single group (groups N2, N3 and N4). The AUC values are shown in Table 1. LipL32 L. interrogans Copenhageni provided best accuracy (AUC = 0.869), followed by LigB [131-649aa] L. kirschneri Gryppotyphosa (AUC = 0.855) and LigB [131-649aa] L. interrogans Canicola (AUC = 0.826). Furthermore, when group P1 was compared to groups N2, N3 and N4 separately, the same antigens showed best performance. The combination of those three antigens improved the diagnostic accuracy when considering all control samples in a single group (Figure 3B) and also when group P1 was compared to group N3 (stray dogs, Figure 4). Using group N2 as the control group, LipL32 and LigB [131-649aa] L. kirschneri Gryppotyphosa provided the best performance. When we consider group N4 (dogs with other febrile syndromes) as control, however, the addition of extra antigens did not impact the diagnostic performance of LipL32 (Figure 4).Figure 3

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