Diagnosis of Indian Visceral Leishmaniasis by Nucleic Acid Detection Using PCR

作者：Srivastava Pankaj^*; Mehrotra Sanjana; Tiwary Puja; Chakravarty Jaya; Sundar Shyam

来源：PLos One, 2011, 6(4): e19304.

DOI：10.1371/journal.pone.0019304

Background: PCR based diagnosis for Visceral Leishmaniasis (VL), despite numerous published primers, remains far from being applied in the field. The present study was planned to design a Leishmania specific diagnostic assay and to evaluate its sensitivity and specificity on a sample size, which to the best of our knowledge is the largest ever screened in one study.
Methods: Leishmania specific primers were developed using 18S rRNA gene and their sensitivity was evaluated on 500 parasitologically confirmed patients with VL and 25 Post Kala-azar Dermal Leishmaniasis (PKDL) patients. Specificity was calculated on 250 healthy endemic controls, 250 healthy non endemic controls and 250 non leishmanial diseases like malaria.
Results: Our PCR assay had a sensitivity of 87.8% (95% CI: 84.1-89.8) using 200 mu L of patient's peripheral-blood. Specificity was absolute in non-endemic healthy controls and in subjects with different diseases while in endemic controls it was 84% (95% CI: 78.9-88.0). Its overall specificity was 94.6% (95% CI-92.8-96.1).
Conclusions: The PCR assay developed is sensitive enough to detect the 18S rRNA gene in an amount equivalent to a single parasite or less in a one million human cell environment. The high sensitivity of this PCR diagnostic test with relatively non-invasive peripheral blood sampling method opens up the possibility of its deployment in field for the routine diagnosis of VL.

• 出版日期2011-4-29
• 单位河北医科大学