Molecular characterization of Babesia microti seroreactive antigen 5-1-1 and development of rapid detection methods for anti-B. microti antibodies in serum

Cai, Yangchuan; Wu, Fen; Hu, Wei; Chen, Jiaxu; Chen, Shao Hong; Xu, Bin; Lü, Yan; Ai, Lin; Yang, Chun li; Zhao, Shimin

doi:10.1016/j.actatropica.2018.03.020

Cited by 5 publications

(14 citation statements)

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“…BALB/c mice were purchased from the Institute of Zoology, Chinese Academy of Sciences, and maintained at the animal care center of the National Institute of Parasitic Diseases, China CDC (IPD, China CDC). The B. microti Peabody strain (ATCC, PRA-99) was obtained from the American Type Culture Collection (Manassas, VA) and maintained in BALB/c mice by serial passage, following the method described previously ( 49 ).…”

Section: Methodsmentioning

confidence: 99%

“…Western blot analysis was performed using standard methods, as described previously ( 49 ). Briefly, proteins extracted from iRBCs at 8 days post-infection and from uninfected RBCs, as well as the purified rN-BmRON2 and rC-BmRON2, were fractioned by SDS-PAGE.…”

Section: Methodsmentioning

confidence: 99%

“…Indirect immunofluorescent assays (IFA) were performed to localize the native BmRON2 within B. microti parasites, as described previously ( 49 ). Briefly, infected red blood cells were fixed with 100% methanol on microscope slides.…”

Section: Methodsmentioning

confidence: 99%

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Molecular Characterization and Immunological Evaluation of Truncated Babesia microti Rhoptry Neck Protein 2 as a Vaccine Candidate

Cai

Yang

et al. 2021

Front. Immunol.

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Babesia microti is a protozoan that infects red blood cells. Babesiosis is becoming a new global threat impacting human health. Rhoptry neck proteins (RONs) are proteins located at the neck of the rhoptry and studies indicate that these proteins play an important role in the process of red blood cell invasion. In the present study, we report on the bioinformatic analysis, cloning, and recombinant gene expression of two truncated rhoptry neck proteins 2 (BmRON2), as well as their potential for incorporation in a candidate vaccine for babesiosis. Western blot and immunofluorescence antibody (IFA) assays were performed to detect the presence of specific antibodies against BmRON2 in infected mice and the localization of N-BmRON2 in B. microti parasites. In vitro experiments were carried out to investigate the role of BmRON2 proteins during the B. microti invasion process and in vivo experiments to investigate immunoprotection. Homologous sequence alignment and molecular phylogenetic analysis indicated that BmRON2 showed similarities with RON2 proteins of other Babesia species. We expressed the truncated N-terminal (33–336 aa, designated rN-BmRON2) and C-terminal (915–1171 aa, designated rC-BmRON2) fragments of the BmRON2 protein, with molecular weights of 70 and 29 kDa, respectively. Western blot assays showed that the native BmRON2 protein is approximately 170 kDa, and that rN-BmRON2 was recognized by serum of mice experimentally infected with B. microti. Immunofluorescence analysis indicated that the BmRON2 protein was located at the apical end of merozoites, at the opposite end of the nucleus. In vitro red blood cell invasion inhibition studies with B. microti rBmRON2 proteins showed that relative invasion rate of rN-BmRON2 and rC-BmRON2 group is 45 and 56%, respectively. Analysis of the host immune response after immunization and B. microti infection showed that both rN-BmRON2 and rC-BmRON2 enhanced the immune response, but that rN-BmRON2 conferred better protection than rC–BmRON2. In conclusion, our results indicate that truncated rhoptry neck protein 2, especially its N-terminal fragment (rN-BmRON2), plays an important role in the invasion of host red blood cells, confers immune protection, and shows good potential as a candidate vaccine against babesiosis.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Molecular Characterization and Immunological Evaluation of Truncated Babesia microti Rhoptry Neck Protein 2 as a Vaccine Candidate

Cai

Yang

et al. 2021

Front. Immunol.

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“…microti antibodies. Therefore, when these parasites infect a new host again, they can promote the production of higher antibody levels (Cai et al, 2018).…”

Section: Discussionmentioning

confidence: 99%

Transmission risk evaluation of transfusion blood containing low-density Babesia microti

Cai,

Xu,

Liu

et al. 2024

Front. Cell. Infect. Microbiol.

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BackgroundBabesia is a unique apicomplexan parasite that specifically invades and proliferates in red blood cells and can be transmitted via blood transfusion, resulting in transfusion-transmitted babesiosis. However, detecting Babesia in blood before transfusion has not received enough attention, and the risk of transfusing blood containing a low density of Babesia microti (B. microti) is unclear, possibly threatening public health and wellness.PurposeThis study aimed to determine the lower detection limit of B. microti in blood and to evaluate the transmission risk of blood transfusion containing low-density B. microti.MethodsInfected BALB/c mouse models were established by transfusing infected whole blood with different infection rates and densities of B. microti. Microscopic examination, nested Polymerase Chain Reaction (nested PCR), and an enzyme-linked immunosorbent assay (ELISA) were used to evaluate the infection status of the mouse models. Meanwhile, the nested PCR detection limit of B. microti was obtained using pure B. microti DNA samples with serial concentrations and whole blood samples with different densities of B. microti-infected red blood cells. Thereafter, whole mouse blood with a B. microti density lower than that of the nested PCR detection limit and human blood samples infected with B. microti were transfused into healthy mice to assess the transmission risk in mouse models. The infection status of these mice was evaluated through microscopic examination, nested PCR tests, and ELISA.ResultsThe mice inoculated with different densities of B. microti reached the peak infection rate on different days. Overall, the higher the blood B. microti density was, the earlier the peak infection rate was reached. The levels of specific antibodies against B. microti in the blood of the infected mice increased sharply during the first 30 days of infection, reaching a peak level at 60 days post-infection, and maintaining a high level thereafter. The nested PCR detection limits of B. microti DNA and parasite density were 3 fg and 5.48 parasites/μL, respectively. The whole blood containing an extremely low density of B. microti and human blood samples infected with B. microti could infect mice, confirming the transmission risk of transfusing blood with low-density B. microti.ConclusionWhole blood containing extremely low density of B. microti poses a high transmission risk when transfused between mice and mice or human and mice, suggesting that Babesia detection should be considered by governments, hospitals, and disease prevention and control centers as a mandatory test before blood donation or transfusion.

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“…are needed. Recently, an immunochromatographic test based on a recombinant B. microti surface antigen showed promising results in experimentally infected mice (Cai et al 2018 ).…”

Section: Relevant Zoonotic Tbds and Commercial Availability Of Diagnostic Test Kits By The Example Of Germanymentioning

confidence: 99%

Tick-borne zoonoses and commonly used diagnostic methods in human and veterinary medicine

et al. 2021

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Around the world, human health and animal health are closely linked in terms of the One Health concept by ticks acting as vectors for zoonotic pathogens. Animals do not only maintain tick cycles but can either be clinically affected by the same tick-borne pathogens as humans and/or play a role as reservoirs or sentinel pathogen hosts. However, the relevance of different tick-borne diseases (TBDs) may vary in human vs. veterinary medicine, which is consequently reflected by the availability of human vs. veterinary diagnostic tests. Yet, as TBDs gain importance in both fields and rare zoonotic pathogens, such as Babesia spp., are increasingly identified as causes of human disease, a One Health approach regarding development of new diagnostic tools may lead to synergistic benefits. This review gives an overview on zoonotic protozoan, bacterial and viral tick-borne pathogens worldwide, discusses commonly used diagnostic techniques for TBDs, and compares commercial availability of diagnostic tests for humans vs. domestic animals, using Germany as an example, with the aim of highlighting existing gaps and opportunities for collaboration in a One Health framework.

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Molecular characterization of Babesia microti seroreactive antigen 5-1-1 and development of rapid detection methods for anti-B. microti antibodies in serum

Cited by 5 publications

References 43 publications

Molecular Characterization and Immunological Evaluation of Truncated Babesia microti Rhoptry Neck Protein 2 as a Vaccine Candidate

Molecular Characterization and Immunological Evaluation of Truncated Babesia microti Rhoptry Neck Protein 2 as a Vaccine Candidate

Transmission risk evaluation of transfusion blood containing low-density Babesia microti

Tick-borne zoonoses and commonly used diagnostic methods in human and veterinary medicine

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