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 clinica...
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| Format: | Article |
| Language: | English |
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BMC
2020-07-01
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| Series: | Parasites & Vectors |
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| Online Access: | http://link.springer.com/article/10.1186/s13071-020-04246-4 |
| _version_ | 1811331504526065664 |
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| author | Jinming Wang Shandian Gao Shangdi Zhang Xin He Junlong Liu Aihong Liu Youquan Li Guangyuan Liu Jianxun Luo Guiquan Guan Hong Yin |
| author_facet | Jinming Wang Shandian Gao Shangdi Zhang Xin He Junlong Liu Aihong Liu Youquan Li Guangyuan Liu Jianxun Luo Guiquan Guan Hong Yin |
| author_sort | Jinming Wang |
| collection | DOAJ |
| description | 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. |
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| institution | Directory Open Access Journal |
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| language | English |
| last_indexed | 2024-04-13T16:21:32Z |
| publishDate | 2020-07-01 |
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| series | Parasites & Vectors |
| spelling | doaj.art-17381cea30ee46afad45ffbc35c0c31a2022-12-22T02:39:55ZengBMCParasites & Vectors1756-33052020-07-011311710.1186/s13071-020-04246-4Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flowJinming Wang0Shandian Gao1Shangdi Zhang2Xin He3Junlong Liu4Aihong Liu5Youquan Li6Guangyuan Liu7Jianxun Luo8Guiquan Guan9Hong Yin10State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceState Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceDepartment of Clinical Laboratory, The Second Hospital of Lanzhou UniversityState Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceState Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceState Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceState Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceState Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceState Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceState Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceState Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural ScienceAbstract 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.http://link.springer.com/article/10.1186/s13071-020-04246-4Ovine babesiosisHuman babesiosisBabesia motasiCross-priming amplificationVertical flow visualization stripDetection |
| spellingShingle | Jinming Wang Shandian Gao Shangdi Zhang Xin He Junlong Liu Aihong Liu Youquan Li Guangyuan Liu Jianxun Luo Guiquan Guan Hong Yin Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flow Parasites & Vectors Ovine babesiosis Human babesiosis Babesia motasi Cross-priming amplification Vertical flow visualization strip Detection |
| title | Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flow |
| title_full | Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flow |
| title_fullStr | Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flow |
| title_full_unstemmed | Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flow |
| title_short | Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flow |
| title_sort | rapid detection of babesia motasi responsible for human babesiosis by cross priming amplification combined with a vertical flow |
| topic | Ovine babesiosis Human babesiosis Babesia motasi Cross-priming amplification Vertical flow visualization strip Detection |
| url | http://link.springer.com/article/10.1186/s13071-020-04246-4 |
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