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Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flow.


ABSTRACT:

Background

Babesia motasi is known as an etiological agent of human and ovine babesiosis. Diagnosis of babesiosis is traditionally performed by microscopy, examining Giemsa-stained thin peripheral blood smears. Rapid detection and accurate identification of species are desirable for clinical care and epidemiological studies.

Methods

An easy to operate molecular method, which requires less capital equipment and incorporates cross-priming amplification combined with a vertical flow (CPA-VF) visualization strip for rapid detection and identification of B. motasi.

Results

The CPA-VF targets the 18S rRNA gene and has a detection limit of 50 fg per reaction; no cross reaction was observed with other piroplasms infective to sheep or Babesia infective to humans. CPA-VF and real-time (RT)-PCR had sensitivities of 95.2% (95% confidence interval, CI 78.1-99.4%) and 90.5% (95% CI 72-97.6%) and specificities of 95.8 (95% CI 80.5-99.5%) and 97.9 (95% CI 83.5-99.9%), respectively, versus microscopy and nested (n) PCR combined with gene sequencing. The clinical performance of the CPA-VF assay was evaluated with field blood samples from sheep (n = 340) in Jintai county, Gansu Province, and clinical specimens (n = 492) obtained from patients bitten by ticks.

Conclusions

Our results indicate that the CPA-VF is a rapid, accurate, nearly instrument-free molecular diagnostic approach for identification of B. motasi. Therefore, it could be an alternative technique for epidemiological investigations and diagnoses of ovine and/or human babesiosis caused by B. motasi, especially in resource-limited regions.

SUBMITTER: Wang J 

PROVIDER: S-EPMC7391542 | biostudies-literature | 2020 Jul

REPOSITORIES: biostudies-literature

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Publications

Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flow.

Wang Jinming J   Gao Shandian S   Zhang Shangdi S   He Xin X   Liu Junlong J   Liu Aihong A   Li Youquan Y   Liu Guangyuan G   Luo Jianxun J   Guan Guiquan G   Yin Hong H  

Parasites & vectors 20200729 1


<h4>Background</h4>Babesia motasi is known as an etiological agent of human and ovine babesiosis. Diagnosis of babesiosis is traditionally performed by microscopy, examining Giemsa-stained thin peripheral blood smears. Rapid detection and accurate identification of species are desirable for clinical care and epidemiological studies.<h4>Methods</h4>An easy to operate molecular method, which requires less capital equipment and incorporates cross-priming amplification combined with a vertical flow  ...[more]

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