|
KMID : 0613820090190091328
|
|
Journal of Life Science
2009 Volume.19 No. 9 p.1328 ~ p.1332
|
|
|
Application of the 18S Ribosomal DNA (rDNA) PCR-RFLP Technique for the Differential Diagnosis of Anisakidosis
|
|
Kim Sun-Mi
Cho Min-Kyoung
Yu Hak-Sun
Cha Hee-Jae
Ock Mee-Sun
|
|
|
Abstract
|
|
|
|
Anisakidosis is caused by anisakid nematodes (family Anisakidae) larvae which can cause not only direct tissue damage but also a severe allergic response related to excretory-secretion products. Lots of different species of anisakid larvae, including Anisakis simplex, Contracaecum, Goezia, Pseudoterranova, and Hysterothylacium, cause the anisakidosis. But it is difficult to diagnosis the species of larvae since the morphologies of larval anisakid nematodes are almost indistinguishable. In order to diagnosis the differential infections of larval anisakid nematodes, polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) of 18S rDNA - was conducted. Three major species of anisakid larvae including A. simplex, C.ontracaecum spp, and Goezia spp. were collected from mackerel (Scomber japonicus), mullet (Mugil cephalus), founder (Paralichthys olivaceus), eel (Astroconger myriaster) and red sea bream (Pagrus major). PCR amplified 18S rDNA from each species of anisakid larvae was digested with eight restriction enzymes including Taq I, Hinf I, Hha I, Alu I, Dde I, Hae Ⅲ, Sau96 I, and Sau3A I. The original sizes of PCR amplified 18S rDNA were 2.0Kb in both anisakid larvaes and Goezia. Restrction enzymes including Hinf 1, Alu 1, Hha I, Dde 1 and Hae Ⅲ cut differently and distinguished the A. simplex and Contracaecum type C’. However, Contracaecum type A showed two different restriction enzyme cutting patterns by Taq 1, Hinf I, Alu 1, and Dde 1. One of the patterns was the same as those of A. simplex, Contracaecum type C’ and Goezia and the other was unique. These results suggest that PCR-RFLP pattern by Hinf 1, Alu 1, Hae I, Dde 1 and Hae Ⅲ can be applied to differential diagnosis of human infection with A. simplex and Contracaecum type C’. Contracaecum type A needs further study of classification by morphological characteristics and genetic analysis.
|
|
|
KEYWORD
|
|
|
Anisakidosis, Anisakis simplex, Contracaecum, 18S rDNA, PCR-RFLP
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|