Rapid detection of Babesia motasi responsible for human babesiosis by cross-priming amplification combined with a vertical flow.
Animals
Babesia
/ classification
Babesiosis
/ diagnosis
Genes, Protozoan
Humans
Molecular Diagnostic Techniques
/ methods
Nucleic Acid Amplification Techniques
/ methods
Pathology, Molecular
/ methods
RNA, Ribosomal, 18S
/ genetics
Sensitivity and Specificity
Sheep
/ parasitology
Sheep Diseases
/ diagnosis
Babesia motasi
Cross-priming amplification
Detection
Human babesiosis
Identification
Ovine babesiosis
Vertical flow visualization strip
Journal
Parasites & vectors
ISSN: 1756-3305
Titre abrégé: Parasit Vectors
Pays: England
ID NLM: 101462774
Informations de publication
Date de publication:
29 Jul 2020
29 Jul 2020
Historique:
received:
20
01
2020
accepted:
20
07
2020
entrez:
31
7
2020
pubmed:
31
7
2020
medline:
17
3
2021
Statut:
epublish
Résumé
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. 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. 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. 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.
Sections du résumé
BACKGROUND
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
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
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
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.
Identifiants
pubmed: 32727550
doi: 10.1186/s13071-020-04246-4
pii: 10.1186/s13071-020-04246-4
pmc: PMC7391542
doi:
Substances chimiques
RNA, Ribosomal, 18S
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
377Subventions
Organisme : National Basic Research Program of China (973 Program)
ID : 2015CB150300
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