Einzelzellbiologie

Abstrakt

Accentuated Molecular Detection Technique to Segregate and Identify Helminths of Fish through High Resolution Melting (HRM) Analysis

Neeshma Jaiswal , Rashmi Tripathi and Sandeep K Malhotra

Real-time PCR coupled with high-resolution molecular (HRM) analysis was conducted to target the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA. The latter acted as a genetic marker to identify and distinguish two anisakid and one cucullanid species parasitizing marine and freshwater fish. Unique and distinct characteristics of HRM patterns were produced for each of the three roundworms investigated. The melt profiles and threshold of the cycles (Ct values), at which amplification commenced, for Anisakis simplex (Rudolphi), Contrcaecum osculatum (Rudolphi) Dujardin, Contrcaecum sp. and Dacnitoides cotylophora (Ward and Magath) were diagnostic for species. The molecular analyses by sequencing and comparing the internal transcribed spacer (ITS) of the ribosomal DNA of Dacnitoides, Anisakis and Contrcaecum assays established their distinct identity. The present investigation propagates molecular-phylogenetic and morpho-molecular analysis to characterize diagnostic constituents of cucullanid and anisakid roundworms.