Detection and differentiation of ovine Theileria and Babesia by reverse line blotting in China

Niu, Qingli; Luo, Jianxun; Guan, Guiquan; Ma, Miling; Liu, Zhijie; Liu, Aihong; Dang, Zhisheng; Gao, Jinliang; Ren, Qiaoyun; Li, Youquan; Liu, Junlong; Yin, Hong

doi:10.1007/s00436-009-1344-x

Cited by 45 publications

(31 citation statements)

References 29 publications

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“…Interestingly,Shayan et al 2007 could also detect only B. ovis and no B. motasi in Rhipicephalus bursa, Rhipicephalus turanicus, and Rhipicephalus sanguineus. In Turkey, also B. ovis was detected as a pathogenic Babesia species in sheep(Aktas et al 2007;Altay et al 2008;Inci et al 2010) Niu et al (2009). described nine probes as B. motasi in 1,234 blood samples in China.…”

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confidence: 99%

Babesia ovis as the main causative agent of sheep babesiosis in Iran

Ranjbar-Bahadori

Eckert²,

Omidian³

et al. 2011

Parasitol Res

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Babesiosis is a haemoparasitic disease with high economical losses in livestock industry worldwide. The early diagnosis and successful therapy of babesiosis belong to the key steps of control and health management of livestock. Ethanol-fixed blood samples of 400 sheep were analyzed for Babesia infection. Reverse line blot (RLB) was established specifically for Theileria lestoquardi, Theileria (China 1), Theileria (China 2), Theileria ovis, Theileria separata, Babesia ovis, Babesia motasi, Babesia crassa, and Babesia (Lintan). The DNA was extracted from the ethanol-fixed blood samples and amplified with a common primer pair derived from 18S rRNA gene, amplifying both Theileria spp. as well as Babesia spp. Regarding the differences in the length of nucleotide sequences of the polymerase chain reaction (PCR) products obtained from Theileria spp. and Babesia spp., the PCR products derived from Babesia spp. were out screened and analyzed by RLB. The RLB analysis showed that 28 samples within the 400 blood samples were B. ovis positive. No B. motasi, B. crassa, or Babesia (Lintan) could be detected. The sequence analysis of one PCR product as a representative for other B. ovis-positive PCR products confirmed the results of RLB. Our results and the results of other investigators showed that B. ovis could be considered as a main causative agent of sheep babesiosis in Iran. Furthermore, our results also showed that RLB can be used as a reliable method for a simultaneous differentiation of Theileria and Babesia species from each other.

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confidence: 99%

Babesia ovis as the main causative agent of sheep babesiosis in Iran

Ranjbar-Bahadori

Eckert²,

Omidian³

et al. 2011

Parasitol Res

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“…motasi in our study was low (0.4%), consistent with previous data that detected Babesia cf. motasi in field blood samples by RLB assay (Niu et al, 2009). In the present study, only 1 H. longicornis collected from sheep was infected by Babesia cf.…”

Section: Discussionmentioning

confidence: 92%

“…motasi isolates, Babesia sp. Xinjiang, B. major, B. ovata, B. bigemina, B. bovis, T. uilenbergi, T. luwenshuni, T. ovis, T. annulata, T. sinensis, and T. sergenti) containing a N-(trifluoroacetamidohexyl-cyanoethyl, N, N-diisopropyl phosphoramidite [TFA])-C6 amino linker were synthetized (Gubbels et al, 1999;Schnittger et al, 2004;Niu et al, 2009Niu et al, , 2012Abdallah et al, 2016). Positive plasmids from Babesia or Theileria species genomic DNA, 2 species of bacteria (Anaplasma marginale and Borrelia burgdorferi sensu stricto), and water were used as positive, negative, and blank control, respectively, to perform the specificity and sensitivity of the RLB assay (Abdallah et al, 2016).…”

Section: Rlb Hybridizationmentioning

confidence: 99%

“…Varied diagnostic methods have been used for the detection of Theileria and Babesia species in ticks, but the application of molecular methods provides improved sensitivity and specificity and allows direct detection of parasites (Altay et al, 2008;Aydin et al, 2013). Recently, PCR-RLB was used for rapid, simultaneous detection and differentiation of numerous species of Babesia and Theileria even in mixed infection (Schnittger et al, 2004;Niu et al, 2009;Iqbal et al, 2013). Although there have been several studies of the frequency of ixodid tick species and the prevalence of tick-borne diseases in most areas in China (Chen et al, 2014a;Li et al, 2014a;Liu et al, 2015;Yu et al, 2015), little information is available about the prevalence of pathogens in ticks.…”

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confidence: 99%

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Identification of 12 Piroplasms Infecting Ten Tick Species in China Using Reverse Line Blot Hybridization

Abdallah

Niu

Yang

et al. 2017

Journal of Parasitology

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“…Xinjiang and Babesia sp. Lintan [20][21][22][23] . RLB method was developed for differentiation of four Borrelia species in ticks in 1995 24 .…”

Section: Introductionmentioning

confidence: 99%

Description of Reverse Line Blotting (RLB) technique and RLB based studies aimed to determine Theileria and Babesia species in sheep and goats in Turkey

Aydın¹

2016

BSBD

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bsbd@balikesir.edu.tr www.bau-sbdergisi.com ÖZETKeneler tıbbi ve veteriner öneme sahip 200'den fazla patojenin naklinde rol oynamaktadırlar. Bu etkenler omurgalı konakta ve vektör kenelerde tek veya miks enfeksiyonlar şeklinde bulunmaktadır. Mikroskobik ve serolojik yöntemler tür ayrımında karşılaşılan zorluklar ve çapraz reaksiyon gibi dezavantajlara sahiptir. Bu dezavantajların giderilmesi moleküler tanı metotlarının kullanılmaya başlanması ile mümkün olabilmektedir. Son yıllarda, Reverse Line Blotting (RLB) metodu kullanılarak çoklu enfeksiyonların aynı zaman diliminde tespiti yapılabilmektedir. RLB metodu, yeni tür ve genotiplerin keşfedilmesini sağlayan moleküler bir tanı metodudur. RLB testi; cins düzeyinde PZR, türe özgü probların Biodyne-C membrana bağlanması, PZR ürünlerinin denatürasyonu, denatüre ürünlerin problar ile hibridizasyonu, streptavidin-POD + biotin kompleksinin oluşumu, bu kompleksin ışık emisyonunun gerçekleşmesi ve bunun karanlık odada radyografik film üzerine aktarılarak değerlendirilmesi olmak üzere yedi aşamadan oluşmaktadır. Bu derlemede kenelerle bulaşan hastalıkların teşhisinde RLB metodunun kullanımı hakkında detaylı bilgi verilmektedir.Anahtar Kelimeler: Kene, Patojen, PZR, RLB. SUMMARYTicks play a role in transmission of more than 200 pathogens of medical and veterinary importance. These pathogens are present in form of single or mixed infections in the vertebrate host and the vector ticks. Microscopic and serological methods have some disadvantages such as difficulties in species discrimination and cross-reaction problems. Elimination of these disadvantages has been possible by using molecular techniques. Recently reverse line blotting (RLB) method has been used for determination of multiple infections simultaneously. RLB is also useful molecular tool to discover new species and genotypes. RLB is consist of seven steps as the stages of genus level PCR, binding species-specific probes to the Biodyne-C membrane, denaturation of PCR products, hybridization of denatured products with probes, formation of streptavidin-POD + biotin complex, realization of the light emission of this complex and evaluation the result by transferring this on a radiographic film in a dark room. This review provides detailed information about the usage of RLB in the detection of tick-borne infections.

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Detection and differentiation of ovine Theileria and Babesia by reverse line blotting in China

Cited by 45 publications

References 29 publications

Babesia ovis as the main causative agent of sheep babesiosis in Iran

Babesia ovis as the main causative agent of sheep babesiosis in Iran

Identification of 12 Piroplasms Infecting Ten Tick Species in China Using Reverse Line Blot Hybridization

Description of Reverse Line Blotting (RLB) technique and RLB based studies aimed to determine Theileria and Babesia species in sheep and goats in Turkey

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