Prevalence and molecular epidemiology of equine piroplasmosis in China: a neglected tick-borne disease

Chen, Kewei; Hu, Zhe; Li, Jingkun; Wang, Jingfei; Liu, Diqiu; Qi, Ting; Guo, Wanshou; Du, Cheng; Wang, Xiaojun

doi:10.1007/s11427-021-2021-3

Cited by 4 publications

(9 citation statements)

References 6 publications

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“…Numerous assays targeting one or multiple EP parasites, including conventional PCR, nPCR and real‐time PCR methods, have been developed. However, our investigation showed that, the bind region of previously developed real‐time PCR probe has three mutation sites in the Chinese T. equi strain (Bhoora et al., 2018; Chen et al., 2021) (Figure 1a). Additionally, there is no real‐time PCR method for the simultaneous detection and differentiation of T. equi group and B. caballi genotypes prevalent in China.…”

Section: Discussionmentioning

confidence: 81%

“…( S. equi ) strain, which were kept in our laboratory. DNA from T. equi and B. caballi isolates and cDNA/DNA from EHV‐1, EHV‐4, S. abortus equi , S. equi and equine macrophages infected with EIAV were verified firstly by nPCR to be T. equi or B. caballi negative (Chen et al., 2021), and then were used to test specificity of the real‐time PCR assay. The sensitivity of the nPCR and duplex real‐time PCR assays was determined by detecting serially diluted 10‐fold standard plasmids (10 7 ‐10 0 copies/μl).…”

Section: Methodsmentioning

confidence: 99%

“…All DNA samples were evaluated with the nPCR assay, and 18S rRNA gene sequences were amplified from all positive samples. The procedure was the same as described previously (Chen et al., 2021). In brief, primers EP 18S‐1F and EP 18S‐1R (Table 1) were used in the first round of PCR to amplify the nearly full‐length 18S rRNA nucleotide sequence (fragment of approximately 1500 bp).…”

Section: Methodsmentioning

confidence: 99%

“…Moreover, T. haneyi has been identified as genotype C in T. equi group (Bhoora et al, 2020;Knowles et al, 2018). Our previous studies indicated that the T. equi genotypes A, B, C and E are common and widespread in China (Chen et al, 2021).…”

Section: Introductionmentioning

confidence: 99%

“…Real-time PCR methods for detecting EP based on 18S rRNA have been described in previous reports, including single plex real-time PCR assays for T. equi (Kim et al, 2008), and multiplex real-time PCR assays that can detect T. equi and B. caballi (Bhoora et al, 2020;Bhoora et al, 2018;Lobanov et al, 2018). However, compared with the 18S rRNA sequences of Chinese T. equi strains (Chen et al, 2021), there were three loci mutations in the regions identified by probes in the previous studies including sites194 (A to G), 197 (G to T) and 198 (A to G) (Figure 1a). Therefore, a real-time PCR method with a new probe need to be developed, which could detect all the genotypes of T. equi group especially Chinese strains.…”

Section: Introductionmentioning

confidence: 99%

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Development of a duplex real‐time PCR assay for simultaneous detection and differentiation of Theileria equi and Babesia caballi

Chen

Yang

et al. 2022

Transbounding Emerging Dis

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Equine Piroplasmosis (EP) is a tick‐borne disease caused by three apicomplexan protozoan parasites, Theileria equi (T. equi), Babesia caballi (B. caballi) and T. haneyi, which can cause similar clinical symptoms. There are five known 18S rRNA genotypes of T. equi group (including T. haneyi) and three of B. caballi. Real‐time PCR methods for detecting EP based on 18S rRNA analysis have been developed, but these methods cannot detect all genotypes of EP in China, especially genotype A of T. equi. In this study, a duplex real‐time PCR detection method was developed for the simultaneous detection and differentiation of T. equi and B. caballi. The primers and probes for this duplex real‐time PCR assay were designed based on the conserved 18S rRNA gene sequences of all genotypes of T. equi and B. caballi including Chinese strain. Double‐quenched probes were used in this method, which provide less background and more signal to decrease the number of false positives relative to single‐quenched probes. The newly developed real‐time PCR assays exhibited good specificity, sensitivity, repeatability and reproducibility. The real‐time PCR assays were further validated by comparison with a nested PCR assay and a previous developed real‐time PCR for EP and sequencing results in the analysis of 506 clinical samples collected from 2019 to 2020 in eleven provinces and regions of China. Based on clinical performance, the agreements between the duplex real‐time PCR assay and the nPCR assay or the previous developed real‐time PCR assay were 92.5% (T. equi) and 99.4% (B. caballi) or 87.4% (T. equi) and 97.2% (B. caballi). The detection results showed that the positivity rate of T. equi was 43.87% (222/506) (10 genotype A, 1 genotype B, 4 genotype C, 207 genotype E), while that of B. caballi was 5.10% (26/506) (26 genotype A), and the rate of T. equi and B. caballi co‐infection was 2.40% (12/506). The established method could contribute to the accurate diagnosis, pathogenic surveillance and epidemiological investigation of T. equi and B. caballi infections in horses.

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Section: Discussionmentioning

confidence: 81%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Development of a duplex real‐time PCR assay for simultaneous detection and differentiation of Theileria equi and Babesia caballi

Chen

Yang

et al. 2022

Transbounding Emerging Dis

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Molecular detection of piroplasms in domestic donkeys in Xinjiang, China

Cui,

Cao,

et al. 2024

Veterinary Medicine & Sci

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BackgroundPiroplasmosis is a common and prevalent tick‐borne disease that affects equids.ObjectivesTo determine the infection and molecular characteristics of the piroplasms in donkeys from Xinjiang, northwestern China, we undertook a cross sectional study by collecting representative samples across several counties within the region.MethodsA total of 344 blood samples were collected from adult domestic donkeys from 13 counties in Xinjiang. PCR was conducted to test for T. equi and B. caballi in the blood samples based on the equine merozoite antigen‐1 (Ema‐1) gene and the 48 kDa rhoptry protein (BC48) gene, respectively.ResultsSixteen blood samples tested positive for piroplasms and the overall infection rate was 4.7% (16/344). Seven of the 13 counties were positive for piroplasms. Among the 16 piroplasm‐positive samples, 15 were singly infected with T. equi with an infection rate of 4.4% (15/344), and coinfection with T. equi and B. caballi was detected in one sample (0.3%, 1/344) from Wushi. Four T. equi sequence genotypes were identified and grouped into different branches of the evolutionary trees.ConclusionThese findings suggest that the infection rate of piroplasms is low in domestic donkeys in southern Xinjiang and that T. equi genotypes have a regional distribution.

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Diagnostic Performance of Competitive ELISA and Western Blot Methods for the Detection of Antibodies against Theileria equi and Babesia caballi

et al. 2022

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Theileria equi (T. equi) and Babesia caballi (B. caballi) are the causative pathogens of Equine piroplasmosis (EP), a disease that has brought huge economic losses and great restrictions to the global equine industry. Rapid and accurate diagnostic methods are critical for the effective monitoring of the disease. In this study, we developed novel competitive ELISA methods and western blot assays based on the EMA1 or Bc48 proteins to detect antibodies against T. equi or B. caballi, respectively. In the novel cELISA, horseradish peroxidase (HRP)-labeled monoclonal antibodies are used in place of enzyme-conjugated secondary antibodies, in order to speed up the entire procedure. These methods have high sensitivity and no cross-reactivity with antibodies against other equine diseases. In the newly developed western blot assays, we optimized the dilution of T. equi or B. caballi positive serum samples to 1:200. Compared with the commercially available kit, both the novel cELISA assay and the western blot assay showed high coincidence rates in detecting antibodies against T. equi and B. caballi. Taken together, the novel cELISA and the western blot assays for detecting antibodies against T. equi or B. caballi have the potential to rapidly test for T. equi or B. caballi and to contribute to the surveillance and control of this disease.

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Prevalence and molecular epidemiology of equine piroplasmosis in China: a neglected tick-borne disease

Cited by 4 publications

References 6 publications

Development of a duplex real‐time PCR assay for simultaneous detection and differentiation of Theileria equi and Babesia caballi

Development of a duplex real‐time PCR assay for simultaneous detection and differentiation of Theileria equi and Babesia caballi

Molecular detection of piroplasms in domestic donkeys in Xinjiang, China

Diagnostic Performance of Competitive ELISA and Western Blot Methods for the Detection of Antibodies against Theileria equi and Babesia caballi

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