Sequence‐related amplified polymorphism, an effective molecular approach for studying genetic variation in Fasciola spp. of human and animal health significance

Abstract

In the present study, a recently described molecular approach, namely sequence-related amplified polymorphism (SRAP), which preferentially amplifies ORFs, was evaluated for the studies of genetic variation among Fasciola hepatica, Fasciola gigantica and the "intermediate" Fasciola from different host species and geographical locations in mainland China. Five SRAP primer combinations were used to amplify 120 Fasciola samples after ten SRAP primer combinations were evaluated. The number of fragments amplified from Fasciola samples using each primer combination ranged from 12 to 20, with an average of 15 polymorphic bands per primer pair. Fifty-nine main polymorphic bands were observed, ranging in size from 100 to 2000 bp, and SRAP bands specific to F. hepatica or F. gigantica were observed. SRAP fragments common to F. hepatica and the "intermediate" Fasciola, or common to F. gigantica and the "intermediate" Fasciola were identified, excised and confirmed by PCR amplification of genomic DNA using primers designed based on sequences of these SRAP fragments. Based on SRAP profiles, unweighted pair-group method with arithmetic averages clustering algorithm categorized all of the examined representative Fasciola samples into three groups, representing the F. hepatica, the "intermediate" Fasciola, or the F. gigantica. These results demonstrated the usefulness of the SRAP technique for revealing genetic variability between F. hepatica, F. gigantica and the "intermediate" Fasciola, and also provided genomic evidence for the existence of the "intermediate" Fasciola between F. hepatica and F. gigantica. This technique provides an alternative and a useful tool for the genetic characterization and studies of genetic variability in parasites.

Regional Differences In The Amino Acid Sequence Of Fasciola Hepatica Cathepsin L2 From Turkey Isolate

“…populations. Therefore, the accurate analysis of genetic variation in parasites has important implications (20).…”

“…The understanding of the parasite life cycle, transmission, study of population biology, epidemiology, as well as genetic variation in parasite populations is important for the control and effective prevention of parasitic diseases in humans and animals. Thus, the accurate analysis of genetic variation in parasites has important implications (20). Additionally, in order to eliminate or minimize the impact of these parasites and to be used in diagnostic methods and immunization studies, it is also important to understand the genetic diversity of the liver fluke populations and the relationship between parasite and host at regional bases (21).…”

Electrophoretic detection of genetic variability among Schistosoma japonicum isolates by sequence‐related amplified polymorphism

“…The SRAP procedures followed that described by Li and Quiros [22], with slight modifications [24,25]. A total of 10 different combinations of SRAP primers including five forward primers and five reverse primers were tested, and the sequences of these SRAP primers are listed in Table 2.…”

“…Since it was reported by Li and Quiros [22], SRAP has been applied extensively in genetic diversity analysis, comparative genomic and genetic studies in a number of organisms including human and animal parasites [23][24][25].…”

Issues Associated with Genetic Diversity Studies of the Liver Fluke, Fasciola Heptica (Platyhelminthes, Digenea, Fasciolidae)

“…SRAP technique has been found to be useful for revealing genetic variability within and between F. hepatica, F. gigantica and evidential for the existence of the "intermediate Fasciola" from different host species and geographical locations in mainland China (Li et al, 2009). The same technique has been utilized for evaluation of genetic diversity of F. hepatica populations from different host species and 16 geographical locations in Spain (Alasaad et al, 2008).…”