Kenelerle bulaşan hastalıkların Revers Line Blotting (RLB) ile tespiti

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Kenelerle Bulagan Hastahklann

Revers

Line Blotting

(RLB)

ile Tespiti

Mehmet Fatih Aydrn

Karamano$lu Mehmetbey Universitesi, Saghk YUksekokulu, 701 OO, Karaman

veterinerm

6zet

Kenelerin _{ttbbi ve veteriner onemi olan 200'den fazla patojenin naklinde rol aldrgr bilinmektedir.}_{Bu etkenler}

konakta

ve

ara

konak kenelerde

tek

veya miks enfeksiyonlar geklinde bulunmaktadrr. Mikroskobik ve

serolojik test yontemleri uzun sUre bu etkenlerin tespitinde kullanrlmrg fakat mikroskobik yontemler ile tUr

ayrlmtntn yaptlamamast, serolojik yontemler ile de tUrler arasr gapraz reaksiyonlarrn gciriilebilmesi nedeniyle

yeni teknikler kullantlmaya baglanmrgtrr. Polimeraz Zincir Reaksiyonu (PZR)

ile

sadece

bir tur

tespit

edilebilmekte, multiplex PZR ile ise birkag farklt tUr eg zamanlr olarak belirlenebilmektedir. Bununla beraber

kenelerin nakletti$i etkenlerin sayrca

fazla

olmasr aynr

anda

_9ok

sayrda etkenin teghis edilebilmesi

gereklili$ini dogurmugtur. Bu amagla bu etkenlerin tamamrnrn gerek kanda gerekse kenede e9 zamanlr

olarak teghis edilmesine ve yeni

tijr

ve genotiplerin kegfedilmesine olanak saglayan Revers Line Blotting

(RLB) yontemi kullantlmaktadtr. RLB testi cins duzeyinde PZR, tUre ozgu problarrn Biodyne-C membrana

ba$lanmast, PZR Urtinlerinin denatiirasyonu, denature tjrirnlerin problar ile hibridizasyonu, streptavidin-POD

+

biotin kompleksinin olugumu, bu kompleksin rgrk emisyonunun gergeklegmesi ve bunun karanlrk odada

radyografik film iizerine aktartlarak de$erlendirilmesi agamalarrndan olugmaktadlr. Bu derlemede son yrllarda

kullanrmr artan RLB hakkrnda bilgi verilmigtir.

Anahtar Kelimeler: Kene, patojen, PZR, RLB

Detection of Tick-borne

Diseases

w:th

Revers

Line Blotting

(RLB)

Summary

Ticks are known to be involved in transport more than 200 pathogens of medical and veterinary importance.

These agents present in the form

of

single

or

mixed infections in the host and intermediate host ticks.

Microscopic and serological testing methods used

in

determining these agents for

a

long time but new

techniques have been used because

of

the

diffuculties

in

species discrimination with

the

microscopic

methods and cross-reactions inter-species with serological methods.

lt

can be detected only one species

with

polymerase

chain

reaction (PCR)

and

the

multiplex

PCR

can

be

determined

a

few

species

simultaneously. However, because of ticks transport a great number of agents, it is necessary that numerous

agents should be diagnosed at the same time. For this purpose, reverse line blotting (RLB) method which

allows diagnosis

all

these agents

in

blood

or

tick

simultaneously and discovery

of

new species and

genotypes

is

used. RLB consists the stages

of

genus level PCR, binding species-specific probes to the

Biodyne-C membrane, denaturation

of

PCR products, hybridization

of

denatured products with probes,

formation

of

streptavidin-POD

+

biotin complex, realization

of

the

light emission

of

this

complex and

evaluation

the

result

by

transferring this

on

a

radiographic

film

in

a

dark

room. This review provides

information about RLB with the increasing use in recent years.

Keywords: Tick, pathogen, PCR, RLB