Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243 122, India (Received March 16, 2000

!

S. H. B, J. R. R, SwatiO, A. K. T, R. K. S,

R. S. KandG. B

National Biotechnology Centre; Division of Parasitology, Indian Veterinary Research Institute, Izatnagar 243 122, India

(Received March 16, 2000; accepted October 30, 2000)

A highly reproducible, dominant, monomorphic fragment of 473 base pair (bp) amplified from the genomeof by arbitrary primer - polymerase chain reaction (AP-PCR) was labelled with digoxigenin and investi- gated for its potential as DNA probe. Dot-blot hybridisation of total genomic DNA with the probe proved useful in detecting bubaline, cameline and equine strains of down to 10 pg of parasite template DNA. No cross-hybridis-

ation was seen with and the bubaline host

DNA. This probe may facilitate laboratory identification of in develop- ing countries, without the inherent risk associated with radioisotopes.

AP-PCR, , non-isotopic probe

! " " "

# $%%&' ( ) $%%*' + ,- $%%*' ( . $%%* $%%/' . $%%0' 12 . $%%%. # " " " ! ! . 32 "

" " . 4 ! ! "

"!" - 2 .

4 ! ! 2" "

" . 5 " " " "

. 6 ".

" ! 2 2 ! " . $%%/. 6 2 " " ! .

6 " " 2 ! ! " ". 4!

2 " " 7 88

! 3 0.&

"" 9 #" $%:&. 4 2 ; "

5 . $%0%.

*:< " " ! /7!4#4#4!<7 ! ,!

. $%%% 2 " # , # 6. 4 " 2 "

" " ! , . $%%%.

" *:< " " 2 ! " ;!$$!=4 6#!

, # !

"7 . 4 2 /& >

$/ $& 4!3 /& ? $ !6#

; @&& > " 4 44 <&A 2 ; 6#!=4 $/ $./ = ! . 4 " 2 4 B ! (2 =5. 2 , . $%%%. 4 " 2 !

" " < > "

$.*A .

C $ @/

$&& > " &.$ 4!84 3 0.& 2 " / . 4 / > " $& 13 2 " $& . 4 $&/ > " @ 2 + #4 , " ! ! ,. 5 " " @/

" "

2 . 4 2 @ D 55 " E =F !

$@& B^± " / =F =F =

=5." G0 H < 2 G0 H 4 =? !

"##"#$%&'(')*&(+$$%%

"" @/ > " " 6#! . "!

2 2 2 2 @ D 55 &./A 2I 55 " $& "2 2 2 G0 H " $/

&.$A 55 &.$A 2I 55 . 4 ! 2 2 !;! E $J$&&&

$A 2 ,4IK!

C. " " ! " *:< - C. $. 4 " "!

2 6#! ! 2 !

" 2 C. $,. ( " "

"" 2 " $ $&

$&& $ $& @/ " 2 2

$& . 32 2 2 C. $. 4 " ! "

2.

!"#$#$#%" & ' (%

) ' *+ , -%.. *+/000 1 2 2 (% *0$ 2 23

*+ 4 4 4 *+

152 23 *+3 ' 5 ) ' .! ) ' 6 ) ' ) ' 66 ) ' 6 ) '

4 " "

" G@ $&& " 5 ! ! # $%%&' F $%%&' + ,- $%%*.

4 " " "

I "

. 4 " " ! " !

" " " " " 2 " 2 " " ! #E $%%*' . $%%0. 6 " ""

" 2 " " ! . ! ! " "

" " ! " " " !

" " .

12 . $%%%

" ; " . 4 "

" 2 G0 H " 2 ! ; ! 2 " . )

" ! 2 " .

( ;! *:< "!

2 " " " ! # $%%&' F!

$%%&' + ,- $%%*. ! 2 2 ! ! " ! " " ! 6 2 "

2 .

$FNQRZOHGJHPHQWV

The senior author thanks the Director, Indian Veterinary Research Institute (IVRI), Izatnagar for the award of Junior Research Fellowship and for the facilities pro- vided for this work. Thanks are also due to and laboratories, IVRI for providing the and DNA samples.

"##"#$%&'(')*&(+$$%%

5HIHUHQFHV

-54 1 74 4 8 4 14 9 - 4 $ :::3 2 *+ ; .:'.!!

4 8 < 4 74 84 84 8 - =,4 * ::!3 >

*+ 2 ? @%'@@

&4 1 < $24 * $ :(63 0 5 2 * ? !.'!%

< 4 8 < $A4 ::3 1 *+

2 2 2 ; .@('.((

<4 * 9 $4 = ::63 1

, *+ < - .(:'.B

C4 14 4 8 4 -54 1 74 14 9 - 4 $ :::3 * 5 2 2 ; 7 %!'%!:

4 8 &4 4 4 <4 +4 -4 $ 14 C ::B3 5 2 *+ 2 ; '@

24 D 4 4 8 4 - 4 $ 14 - ::B3 2

*+ 2 2 2 ; %!'%.%

1,4 84 E 4 E <4 # :B:3 < 3 &2 <

1 7 &2 4 1 74 + D,

;,4 + 24 1 ::63 1 *+ 5 ! 1 8 # < 7 .'.(

=4 8 + D4 8 ::3 ,22 5 2 ; ('!@

=24 +4 ,AA4 + 24 1 ::3 2 5 2 *+ 1 8 # <

7 .@@'.(

=24 +4 ,AA4 +4 14 + 24 1 ::!3 2 - 1 # < ..:'.%(

F4 F G -H4 # ::3 0 "#

5 *+ ; :('.6B