Comparative evaluation of the sensitivity of LAMP, PCR and in vitro culture methods for the diagnosis of equine piroplasmosis
Comparative evaluation of the sensitivity of LAMP, PCR and in vitro culture methods for the diagnosis of equine piroplasmosis
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DOI：
10.1007/s00436-006-0430-6
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发表时间：
2007-04-01
影响因子：
2
通讯作者：
Igarashi, Ikuo
中科院分区：
文献类型：
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作者：
Alhassan, Andy;Govind, Yadav;Igarashi, Ikuo
采用10倍稀释培养的马泰勒虫和卡巴里虫，评价了LAMP、PCR和体外培养方法检测马泰勒虫和卡巴里虫的敏感性。在培养后第1天，T equi和B.在体外培养方法中，仅在1%寄生虫稀释度下可观察到caballi寄生虫，而LAMP可检测高达1 × 10（-3）%的马锥虫和B。而PCR可检测1 × 10（-3）%马锥虫和1 × 10（-1）%B。卡巴利寄生虫稀释液。在培养后第7天，马类泰勒虫和B.在体外培养中，卡氏锥虫的检出限增加到1 × 10（-6）%，而LAMP对两种寄生虫的检出限增加到1 × 10（-10）%，而PCR对马锥虫和B的检出限增加到1 × 10（-10）%和1 × 10（-6）%。caballi，分别。此外，LAMP和PCR扩增从实验感染的马的器官中提取的马绦虫DNA。本研究进一步验证了LAMP作为一种替代的分子诊断工具，可用于马梨形虫病的早期诊断，也可与PCR一起作为死后检验的辅助方法。
The sensitivity of LAMP, PCR and in vitro culture methods for the detection of Theileria equi and Babesia caballi was evaluated using tenfold serially diluted culture parasites. On day 1 post-culture, both T equi and B. caballi parasites could only be observed at 1% parasite dilution from the in vitro culture method, whereas LAMP could detect up to I X 10(-3)% of both T equi and B. caballi parasite dilutions, whilst PCR could detect I X 10(-3)% T equi and I x 10(-1)% B. caballi parasite dilutions. On day 7 post-culture, the detection limit for T equi and B. caballi in the in vitro culture increased up to I X 10(-6)%, whereas LAMP detection limit increased to I x 10(-10)% for both parasites, whilst the PCR detection limit increased to I x 10(-10)% and I X 10(-6)% for T equi and B. caballi, respectively. Furthermore, LAMP and PCR amplified the T equi DNA extracted from the organs of an experimentally infected horse. This study further validates LAMP as an alternative molecular diagnostic tool, which can be used in the diagnosis of early infections of equine piroplasmosis and together with PCR can also be used as supplementary methods during post-mortems.