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. 2022 Feb 22:12:835383.
doi: 10.3389/fcimb.2022.835383. eCollection 2022.

Chagas Immunochromatographic Rapid Test in the Serological Diagnosis of Trypanosoma cruzi Infection in Wild and Domestic Canids

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Chagas Immunochromatographic Rapid Test in the Serological Diagnosis of Trypanosoma cruzi Infection in Wild and Domestic Canids

Esthefany S Rodrigues et al. Front Cell Infect Microbiol. .

Abstract

Canis lupus familiaris (domestic dog) represents a reliable sentinel for the occurrence of a well-established transmission cycle of Trypanosoma cruzi among wild mammals in the surroundings and, consequently, where the risk of human infection exists. Serological diagnosis is the chosen method to identify T. cruzi infection in dogs that, in Brazil, rarely present positive parasitological tests. The use of recombinant chimeric parasitic antigens results in a sensitive and specific serological diagnostic test in contrast to the use of crude T. cruzi antigens. Our objective was to evaluate the Chagas/Bio-Manguinhos Lateral Flow Immunochromatographic Rapid Test (Chagas-LFRT) for the diagnosis of T. cruzi infection in domestic dogs and the potential of application of this diagnostic platform to wild canid species. Two recombinant proteins (IBMP-8.1 and IBMP-8.4) that displayed the best performance in the enzyme immunoassay (ELISA) in previous studies were tested in a platform with two diagnostic bands. A panel of 281 dog serum samples was evaluated: 133 positive for T. cruzi by serological diagnosis, including 20 samples with positive blood cultures belonging to different discrete typing units (DTUs); 129 negative samples; and 19 samples from dogs infected by other trypanosomatids: Leishmania infantum, Trypanosoma rangeli, Trypanosoma caninum and Crithidia mellificae, in addition to samples infected by Anaplasma platys, Dirofilaria immitis and Erlichia sp. that were employed to evaluate eventual cross-reactions. We also evaluated the Chagas-LFRT to detect T. cruzi infection in 9 serum samples from six wild canid species. We observed that the intensity pattern of the bands was directly proportional to the serological titer observed in IFAT. The sensitivity was 94%, the specificity was 91% according to the ROC curve, and the defined cutoff was an optical density of 4.8. The agreement obtained was considered substantial by the kappa analysis (84%). From T. cruzi positive hemoculture samples, 88.9% were positive by Chagas-LFRT. The test was efficient in recognizing infections by five of the six T. cruzi DTUs. Cross-reactions were not observed in infections by L. infantum, T. rangeli, T. caninum and D. immitis; however, they were observed in sera of dogs infected by Crithidia mellificae, Anaplasma sp. and Erlichia sp. A strong reaction was observed when serum samples from wild canids were submitted to the Protein A affinity test, confirming its applicability for these species. This test will allow rapid preventive actions in areas with high risk to the emergence of Chagas disease in a safer, reliable, low-cost and immediate manner, without the need for more complex laboratory tests.

Keywords: Trypanosoma cruzi; dead-end and sentinel hosts; domestic and wild canids; quimeric antigens; rapid test.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Scheme showing the serum samples from wild and domestic canids with previous serological diagnosis (IFAT and ELISA) selected to compose the panel of positive and negative samples and serum samples with different parasitic infections used in the Chagas lateral flow immunochromatografic test (Chagas-LFRT).
Figure 2
Figure 2
(A) Lateral flow assay (LFA) device. (a) Schematic illustration of the composition of the LFA strip for the detection of IgG anti-Trypanosoma cruzi in samples employing the IBMP-8.4 and IBMP-8.1 chimeric antigens and photographic image of an LFA opened device after loading T. cruzi-specific IgG antibodies. (b) Schemes of the results expected for valid and invalid results yielded by LFA. Copyright© Silva et al., 2020, https://doi.org/10.1155/2020/1803515. (B) Canis lupus familiaris: LBT 2167 (-), LBT 2828 (+), LBT 844 (+; DTU TcI), LBT 1819 (+; DTU TcI/TcII), LBT 6476 (+; DTU TcIII), LBT 8600 (+; DTU TcIII/V), LBT 12079 (+; DTU TcIV); Canis lupus: LBT 4838 (+); LBT 8599: sample from a domestic dog with seroconversion event (2009 and 2016).
Figure 3
Figure 3
BoxPlot with the distribution of mean values of optical densities and mean values of negative and positive samples from dogs infected by Trypanosoma cruzi against the chimeric antigens IBMP-8.1 and IBMP-8.4.
Figure 4
Figure 4
Mean of negative and positive samples from dogs infected by Trypanosoma cruzi against the chimeric antigens IBMP-8.1 and IBMP-8.4. Number of samples (n) and mean optical densities obtained by serological titer (A - negative, B - 1/10, C - 1/20, D - 1/40, E - 1/80, F - 1/160, G - 1/320, H - 1/640, I - 1/1280, J - 1/2560, K – 1/5120, L – 1/10240).
Figure 5
Figure 5
(A) Receiver operating characteristic (ROC) curve and area under the curve (AUC) of cutoff point value 4.8 and (B) AUC of cutoff (-) >1/20 cutoff (-) >1/40.

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