A pan-Theileria FRET-qPCR survey for Theileria spp. in ruminants from nine provinces of China

Particle and Fibre Toxicology

Chengming Wang¹  Heba S El-Mahallawy⁴  Jing Li¹  Jilei Zhang¹  Lu Luan¹  Zhangpin Yang³  Patrick Kelly²  Yongjiang Mao³  Yi Yang¹ 

[1] Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University College of Veterinary Medicine, Yangzhou 225009, Jiangsu, China;Ross University School of Veterinary Medicine, Basseterre 00265, St. Kitts & Nevis;Yangzhou University College of Animal Science and Technology, Yangzhou 225009, Jiangsu, China;Faculty of Veterinary Medicine, Suez Canal University, Ismailia 41225, Egypt

关键词: Ruminants;    Prevalence;    FRET-qPCR;    Theileria spp;   

Others  :  1150541
DOI  :  10.1186/1756-3305-7-413

 received in 2014-07-03, accepted in 2014-08-24,  发布年份 2014

【 摘 要 】

Background

Theileria spp. are tick transmitted protozoa that can infect large and small ruminants causing disease and economic losses. Diagnosis of infections is often challenging, as parasites can be difficult to detect and identify microscopically and serology is unreliable. While there are PCR assays which can identify certain Theileria spp., there is no one PCR that has been designed to identify all recognized species that occur in ruminants and which will greatly simplify the laboratory diagnoses of infections.

Methods

Primers and probes for a genus-specific pan-Theileria FRET-qPCR were selected by comparing sequences of recognized Theileria spp. in GenBank and the test validated using reference organisms. The assay was also tested on whole blood samples from large and small ruminants from nine provinces in China.

Results

The pan-Theileria FRET-qPCR detected all recognized species but none of the closely related protozoa. In whole blood samples from animals in China, Theileria spp. DNA was detected in 53.2% of the sheep tested (59/111), 44.4% of the goats (120/270) and 30.8% of the cattle (380/1,235). Water buffaloes (n = 29) were negative. Sequencing of some of the PCR products showed cattle in China were infected with T. orientalis/T. sergenti/T. buffeli group while T. ovis and T. luwenshuni were found in sheep and T. luwenshuni in goats. The prevalence of Theileria DNA was significantly higher in Bos p. indicus than in Bos p. taurus (77.7% vs. 18.3%) and copy numbers were also significantly higher (10^4.88 vs. 10^3.00Theileria 18S rRNA gene copies/per ml whole blood).

Conclusions

The pan-Theileria FRET-qPCR can detect all recognized Theileria spp. of ruminants in a single reaction. Large and small ruminants in China are commonly infected with a variety of Theileria spp.

【 授权许可】

   
2014 Yang et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

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