Loop-mediated isothermal amplification (LAMP) method based on two species-specific primer sets for the rapid identification of Chinese Babesia bovis and B. bigemina

Liu, Aihong; Guan, Guiquan; Du, Pengfei; Gou, Huitian; Liu, Zhijie; Liu, Junlong; Ma, Miling; Yang, Jifei; Li, Youquan; Niu, Qinli; Ren, Qiaoyun; Bai, Quan; Yin, Hong; Luo, Jianxun

doi:10.1016/j.parint.2012.07.004

Cited by 29 publications

(28 citation statements)

References 14 publications

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“…With these considerations, a LAMP assay for detection of H. meleagridis was developed in this study. To our knowledge, this is the first time that the LAMP assay was developed for detection of this protozoan, although LAMP has been used widely in the detection of some other protozoa (Liu et al, 2012;Oyhenart et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

“…E-mail: xmjsc@sina.com.cn (Received 23 August 2013; accepted 16 November 2013) applied successfully for the detection and diagnosis of many viral, bacterial and fungal diseases (Mori et al, 2013). Recently, LAMP assay has also been used successfully in the diagnosis of a number of parasitic diseases such as bovine babesiosis, trichinellosis, bovine trichomonosis, malaria and giardiasis (Liu et al, 2012;Lin et al, 2013;Oyhenart et al, 2013;Surabattula et al, 2013). However, no information on LAMP assay for detection H. meleagridis has been available.…”

Section: Introductionmentioning

confidence: 99%

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Loop-mediated isothermal amplification assay for detection ofHistomonas meleagridisinfection in chickens targeting the 18S rRNA sequences

Tao

2014

Avian Pathology

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Histomonas meleagridis is the causative agent of histomonosis, a disease of gallinaceous fowl characterized by necrotic typhlitis, hepatitis, and high mortality. To develop a rapid and sensitive method for specific detection of H. meleagridis, an assay based on loop-mediated isothermal amplification (LAMP) targeting the 18S rRNA gene was established. The detection limit of the LAMP assay was 10 copies for standard plasmids containing an 18S rRNA gene fragment, which was superior to that of a classical PCR method. Specificity tests revealed that there was no cross-reaction with other protozoa such as Trichomonas gallinae, Blastocytis sp, Tetratrichomonas gallinarum, Plasmodium gallinaceum, Toxoplasma gondii, Eimeria tenella, Leucocytozoon caulleryi and Leucocytozoon sabrazesi. The assay was evaluated for its diagnostic utility using liver and caeca samples collected from suspected field cases, the detection rate was 100 and 97.92%, respectively. These results indicate that the LAMP assay may be a useful tool for rapid detection and identification of H. meleagridis in poultry.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Loop-mediated isothermal amplification assay for detection ofHistomonas meleagridisinfection in chickens targeting the 18S rRNA sequences

Tao

2014

Avian Pathology

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“…One calf was infected with B. bigemina Kunming strain, and another calf was infected with B. bovis Shanxian strain. Both babesia strain were previously collected from Henan and Yunnan province, respectively, and preserved iliquid nitrogen as a gold standard positive control (Liu et al 2012). Daily blood smears were obtained post-infection for the presence of parasites.…”

Section: Animal Infectionmentioning

confidence: 99%

A PCR method targeting internal transcribed spacers: the simultaneous detection of Babesia bigemina and Babesia bovis in cattle

Liu

Guan

Liu

et al. 2014

Acta Parasitologica

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In this study, two pairs of oligonucleotide primers were designed according to the nucleotide sequence of the internal transcribed spacers (ITSs) of Babesia bigemina and B. bovis isolates from China. The primers were used in a multiplex PCR to detect parasite DNA in blood samples from cattle. There was no cross reactions with B. ovata, B. major, B. sp. Kashi, Theileria annulata, T. sergenti, T. sinensis or normal bovine DNA. The sensitivity of multiplex PCR assay was 1 pg and 10 pg DNA for B. bigemina and B. bovis, respectively. A total of 260 field blood samples collected from cattle in five provinces of China were analyzed by multiplex PCR and light microscopy. PCR testing revealed that 7.3% (19/260) and 5.8% (15/260) of cattle were positive for B. bigemina and B. bovis and 1.2% (3/260) of cattle were co-infected with B. bigemina and B. bovis. Using light microscopy, 2.3% (6/260) and 1.5% (4/260) of cattle were infected by B. bigemina and B. bovis, respectively, and no co-infection was found. The results showed that the multiplex PCR developed in the present study could be an alternative diagnostic tool for the detection of B. bovis and B. bigemina infection in cattle.

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“…Furthermore, there is no need of any expensive and complicated laboratory equipment for LAMP assays. All the reaction process can be performed by using a simple heating block or a water bath and the amplification can be directly visualized by using SYBR Green I under ultraviolet light (Liu et al 2012 (Ikadai et al 2004;He et al 2009) but its sensitivity has not been evaluated for the detection of B. felis in cats. This study is the first attempt to optimize LAMP PCR for feline babesiosis caused by B. felis.…”

Section: Introductionmentioning

confidence: 99%

First report on optimization of loop-mediated isothermal amplification (LAMP) for the diagnosis of Babesia felis

Salim

Akhtar

Lateef

et al. 2017

IJAR

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The objective of present study was to optimize loop mediated isothermal amplification (LAMP) assay for the diagnosis of Babesia felis in cats. LAMP primers were designed recognizing four sections of 18SribosomalRNA (18S rRNA) gene of B. felis. The blood samples of cats microscopically positive for Babesia felis were further used to extract deoxyribo neuclic acid (DNA) and the reaction mixture of 25 µL was standardized at 63°C temperature for 1 hour. LAMP assay provided more positive samples than conventional polymerase chain reaction (PCR). The prevalence of B. felis was also determined in cats using this optimized LAMP assay and it was found that the prevalence was more in younger cats as compare to adults. The application of LAMP can be helpful in rapid, reliable and cost effective diagnosis of B. felis in field.

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Loop-mediated isothermal amplification (LAMP) method based on two species-specific primer sets for the rapid identification of Chinese Babesia bovis and B. bigemina

Cited by 29 publications

References 14 publications

Loop-mediated isothermal amplification assay for detection ofHistomonas meleagridisinfection in chickens targeting the 18S rRNA sequences

Loop-mediated isothermal amplification assay for detection ofHistomonas meleagridisinfection in chickens targeting the 18S rRNA sequences

A PCR method targeting internal transcribed spacers: the simultaneous detection of Babesia bigemina and Babesia bovis in cattle

First report on optimization of loop-mediated isothermal amplification (LAMP) for the diagnosis of Babesia felis

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