Department of Tropical Medicine, School of Public Health and Tropical Medicine, Tulane University, New Orleans, Louisiana, United States of America.
Vector-Borne and Infectious Disease Research Center, Tulane University, New Orleans, Louisiana, United States of America.
PLoS Negl Trop Dis. 2020 Dec 17;14(12):e0008932. doi: 10.1371/journal.pntd.0008932. eCollection 2020 Dec.
Chagas disease is a neglected zoonosis of growing concern in the southern US, caused by the parasite Trypanosoma cruzi. We genotyped parasites in a large cohort of PCR positive dogs to shed light on parasite transmission cycles and assess potential relationships between parasite diversity and serological test performance.
METHODOLOGY/PRINCIPAL FINDINGS: We used a metabarcoding approach based on deep sequencing of T. cruzi mini-exon marker to assess parasite diversity. Phylogenetic analysis of 178 sequences from 40 dogs confirmed the presence of T. cruzi discrete typing unit (DTU) TcI and TcIV, as well as TcII, TcV and TcVI for the first time in US dogs. Infections with multiple DTUs occurred in 38% of the dogs. These data indicate a greater genetic diversity of T. cruzi than previously detected in the US. Comparison of T. cruzi sequence diversity indicated that highly similar T. cruzi strains from these DTUs circulate in hosts and vectors in Louisiana, indicating that they are involved in a shared T. cruzi parasite transmission cycle. However, TcIV and TcV were sampled more frequently in vectors, while TcII and TcVI were sampled more frequently in dogs.
CONCLUSIONS/SIGNIFICANCE: These observations point to ecological host-fitting being a dominant mechanism involved in the diversification of T. cruzi-host associations. Dogs with negative, discordant or confirmed positive T. cruzi serology harbored TcI parasites with different mini-exon sequences, which strongly supports the hypothesis that parasite genetic diversity is a key factor affecting serological test performance. Thus, the identification of conserved parasite antigens should be a high priority for the improvement of current serological tests.
恰加斯病是一种被忽视的人畜共患寄生虫病,在美国南部地区日益受到关注,由寄生虫克氏锥虫引起。我们对大量 PCR 阳性犬中的寄生虫进行基因分型,以揭示寄生虫传播周期,并评估寄生虫多样性与血清学检测性能之间的潜在关系。
方法/主要发现:我们使用基于深度测序的克氏锥虫小外显子标记物的宏条形码方法来评估寄生虫多样性。对来自 40 只犬的 178 个序列的系统发育分析证实了 T. cruzi 离散型单元(DTU)TcI 和 TcIV 的存在,以及在美国犬中首次发现 TcII、TcV 和 TcVI。38%的犬存在多重 DTU 感染。这些数据表明,美国犬中的 T. cruzi 遗传多样性比以前检测到的更高。对 T. cruzi 序列多样性的比较表明,这些 DTU 中来自宿主和媒介的高度相似的 T. cruzi 株在路易斯安那州循环,表明它们参与了共享的 T. cruzi 寄生虫传播周期。然而,TcIV 和 TcV 在媒介中采样更频繁,而 TcII 和 TcVI 在犬中采样更频繁。
结论/意义:这些观察结果表明,生态宿主适应是涉及 T. cruzi-宿主关联多样化的主要机制。血清学检测呈阴性、不一致或阳性的犬携带不同的小外显子序列的 TcI 寄生虫,这强烈支持寄生虫遗传多样性是影响血清学检测性能的关键因素的假设。因此,鉴定保守的寄生虫抗原应该是改进现有血清学检测的首要任务。