Screening for biomarkers reflecting the progression of Babesia microti infection

Autor: Xiao-Nong Zhou, Xiu-Feng Liu, Xue-Nian Xu, Bin Xu, Yan Zhou, Qunfeng Wu, Xia Zhou, Yu-Chun Cai, Chengsong Sun, Wei Hu, Shen-Bo Chen, Cheng Xunjia, Ting Zhang, Jia-Xu Chen, Jian Li, Yong-Feng Fu, Ruixiang Zhang, Jun-Hu Chen, Ji-Lei Huang
Rok vydání: 2018
Předmět:
Proteomics
0301 basic medicine
Erythrocytes
Screening biomarker
Antibodies
Protozoan
Parasitemia
law.invention
Mice
0302 clinical medicine
Strain ATCC®PRA-99TM
law
Blood plasma
Polymerase chain reaction
Babesiosis
Mice model
Recombinant Proteins
Data Accuracy
Infectious Diseases
Disease Progression
Female
Antibody
030231 tropical medicine
Protein Array Analysis
Protein microarray
Antigens
Protozoan
Enzyme-Linked Immunosorbent Assay
Biology
Babesia microti
Sensitivity and Specificity
lcsh:Infectious and parasitic diseases
03 medical and health sciences
Immune system
Antigen
medicine
Animals
Humans
lcsh:RC109-216
Cell free expression system
Research
medicine.disease
biology.organism_classification
Virology
Disease Models
Animal
030104 developmental biology
Parasitology
Immunoglobulin G
Babesia
biology.protein
Expression and evaluation of proteins
Biomarkers
Zdroj: Parasites & Vectors, Vol 11, Iss 1, Pp 1-15 (2018)
Parasites & Vectors
ISSN: 1756-3305
DOI: 10.1186/s13071-018-2951-0
Popis: Background Babesiosis is caused by the invasion of erythrocytes by parasites of the Babesia spp. Babesia microti is one of the primary causative agents of human babesiosis. To better understand the status of the disease, discovering key biomarkers of the different infection stages is crucial. Results This study investigated B. microti infection in the mouse model from 0 to 270 days post-infection (dpi), using blood smears, PCR assays and ELISA. PCR assays showed a higher sensitivity when compared to microscopic examination. Specific IgG antibodies could be detected from 7 days to 270 dpi. Two-dimensional electrophoresis was combined with western blotting and mass spectrometric analysis to screen for specific reactive antigens during both the peak parasitaemia period (7 dpi) and IgG antibody response peak period (30 dpi) by the infected mice plasma. The 87 positive reactive proteins were identified and then expressed with the wheat germ cell-free system. Protein microarrays of all 87 targeted proteins were produced and hybridized with the serial plasma of infected mice model. Based on the antigen reaction profile during the infection procedure, 6 antigens were selected and expressed in Escherichia coli. Due to an early response to IgM, lower immunoreactivity levels of IgG after two months and higher immunoreactivity level IgG during nine months, four recombinant proteins were selected for further characterization, namely rBm2D97(CCF75281.1), rBm2D33(CCF74637.1), rBm2D41(CCF75408.1) and rBm7(CCF73510.1). The diagnostic efficacy of the four recombinant protein candidates was evaluated in a clinical setting using babesiosis patient plasma. The rBm2D33 showed the highest sensitivity with a positive rate of 62.5%. Additional characterization of the two candidate proteins using a mouse vaccination assay, demonstrated that rBm2D41 could reduce peak parasitaemia by 37.4%, indicating its efficacy in preventing severe babesiosis. Conclusions The detection technologies of microscopic examination, PCR assays and antibody tests showed different sensitivities and accuracy during the different stages of B. microti infection. Antibody detection has a unique significance for B. microti infection in the asymptomatic stages. Using immunoreactivity profiles, biomarkers for disease progression were identified and represent useful information for future the diagnosis and vaccine development for this serious disease of public health significance. Electronic supplementary material The online version of this article (10.1186/s13071-018-2951-0) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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