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Information Journal Paper

Title

DEVELOPMENT AND ASSESSMENT OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) ASSAY FOR THE DIAGNOSIS OF HUMAN VISCERAL LEISHMANIASIS IN IRAN

Pages

  50-59

Abstract

 Background: Parasitological methods for the diagnosis of Visceral leishmania-sis (VL) require invasive procedures, so serological and molecular approaches have been developed but are not generally applicable in the field. We evaluated a loop mediated isothermal amplification (LAMP) assay using blood from VL patients and compared it to NESTED PCR.Methods: Forty-seven subjects with clinical features (fever, hepatosplenomegaly and anemia) were confirmed positive for VL by the direct agglutination test (DAT) at titers>3200. Forty DAT negative individuals from non-endemic areas with no clinical signs or symptoms of VL served as controls. A LAMP assay was performed using a set of six primers targeting Leishmania infantum kinetoplast DNA (kDNA) minicircle gene under isothermal (64 oC) conditions. For NESTED PCR we used primers targeting the kDNA minicircle gene.Results: The LAMP assay provided a detection limit of 1 parasite in 1 ml of PERIPHERAL BLOOD and detected L. infantum DNA in 44 of 47 DAT-confirmed VL cases, with diagnostic sensitivity of 93.6% (95% CI). No L. infantum DNA was amplified in controls, indicating a specificity of 100%. The NESTED PCR yielded sensitivity of 96% (95% CI) and a specificity of 100% (95% CI).Conclusion: The LAMP assay gave results similar to those of NESTED PCR but in a shorter time. The LAMP method is simple; requires no sophisticated equipment; has a short reaction time; and results, indicated by turbidity of the reaction mixture, are observable with the naked eye.

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    APA: Copy

    GHASEMIAN, MEHRDAD, GHARAVI, MOHAMMAD JAVAD, AKHLAGHI, LAME, MOHEBALI, MEHDI, MEAMAR, AHMAD REZA, ARYAN, EHSAN, & OORMAZDI, HORMOZD. (2014). DEVELOPMENT AND ASSESSMENT OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) ASSAY FOR THE DIAGNOSIS OF HUMAN VISCERAL LEISHMANIASIS IN IRAN. IRANIAN JOURNAL OF PARASITOLOGY, 9(1), 50-59. SID. https://sid.ir/paper/304031/en

    Vancouver: Copy

    GHASEMIAN MEHRDAD, GHARAVI MOHAMMAD JAVAD, AKHLAGHI LAME, MOHEBALI MEHDI, MEAMAR AHMAD REZA, ARYAN EHSAN, OORMAZDI HORMOZD. DEVELOPMENT AND ASSESSMENT OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) ASSAY FOR THE DIAGNOSIS OF HUMAN VISCERAL LEISHMANIASIS IN IRAN. IRANIAN JOURNAL OF PARASITOLOGY[Internet]. 2014;9(1):50-59. Available from: https://sid.ir/paper/304031/en

    IEEE: Copy

    MEHRDAD GHASEMIAN, MOHAMMAD JAVAD GHARAVI, LAME AKHLAGHI, MEHDI MOHEBALI, AHMAD REZA MEAMAR, EHSAN ARYAN, and HORMOZD OORMAZDI, “DEVELOPMENT AND ASSESSMENT OF LOOP-MEDIATED ISOTHERMAL AMPLIFICATION (LAMP) ASSAY FOR THE DIAGNOSIS OF HUMAN VISCERAL LEISHMANIASIS IN IRAN,” IRANIAN JOURNAL OF PARASITOLOGY, vol. 9, no. 1, pp. 50–59, 2014, [Online]. Available: https://sid.ir/paper/304031/en

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