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Application of Nested PCR for Confirmative Diagnosis of Canine Ehrlichiosis

Narayana Rao Lenka Shobhamani Bollam Vaikunta Rao Velamala V. K. Subramanyam
Vol 10(4), 85-91

In a retrospective study, 50 Ehrlichia suspected dogs were tested with different diagnostic methods. Among them 8, 18 and 30 percent of samples were detected as positive for ehrlichiosis by peripheral blood smear, buffy coat smear examination and Anigen Rapid E.canis Ab test kit, respectively. The above test results were compared with nested PCR, of them 34 percent samples were positive for E.canis. First step PCR genus-specific primers amplified a 477 bp suggestive band of Ehrlichia and second step species-specific nested PCR amplified at 387 bp suggestive band of Ehrlichia canis. When qualitative results were compared based on Kappa statistical analysis revealed that nested PCR 72% agreement (k= 0.21) with peripheral blood smear examination, 84 % agreement (k=0.59) with buffy coat smear examination and 94% agreement (k=0.86) with Anigen Rapid E.canis Ab test kit. The sensitivity and specificity of the tests were found to be 0.17 (17%) and 1.0 (100%) for the peripheral blood smear, 0.52(52%) and 1.0 (100%) for buffy coat smear examination, and 0.87(87%) and 0.97(97%) for the Anigen Rapid E.canis Ab test kit. In conclusion, nested PCR is more sensitive and reliable diagnostic methods for diagnosis of early as well as subclinical form of canine ehrlichiosis.

Keywords : Anigen Rapid E. canis Ab test kit Blood Smear Examination Ehrlichia canis Nested PCR

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