ISOLATION AND CHARACTERIZATION OF AVIAN LEUKOSIS VIRUS FROM BREEDER FLOCKS OF CHICKEN
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Date
2010-10
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SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA
Abstract
ABSTRACT:
The present study was takcn up with a view to isolate a\.ian Icukc,sis virus
(ALV) from aft'ected hrecdcr flocks of chickens and characterize the isolatc(s)with
rcgard ttr group specitic ac-ELISA, growth in cell culturc titration. serum
neutralization, polymerase chain reaction multiplc scqucnce alignment and
phylogenetic allalysis in diagnosis of avian leucc~sisv irus infection.
276 cloacal swab sarnples wcrc collected fi-on1 hrccder Ilocks 01' chicken
suspcctccl li>r avian lcukosis viral inl'ections. The breedcr flocks of chickcn cxhib~trd
sympto~nsli ke tumours in livcr, splccn and heart. Thc sa~nplcs\s Jcr-ct cstcd tbr ALV
by group spccilic antigen capture El-ISA ;is a prcliniinnry test hcliirc attcnipts t o
isolate the virus. A total of47 sa~nplesw crc positive of'270 samples by cn~pioying
p27 ac-ELISA kit. DNA from 16 blood samples (buffy coat) and RNA from 25
cloacal swabs obtained from ALV gs antigen positive flocks were tested for ALV
specific sequences by PCR Attempts were made to isolate avian leukosis virus from
these cloacal swab samples by passaging in CEF cells. The samples were passaged
five times in cell lines. The presence of virus was demonstrated at different passage
levels by ac-ELISA and Polymerase chain reaction (PCR).
The RT PCR using H5 and AD1 was found negative for the SVVU-I01
isolate where as RT-PCR using primers H5 and H7b was positive with expected
product size of 544 bp, which indicate that SVVU-I01 belongs to ALV subgroup-.I.
Virus neutralization results indicate that the homologous antiserum efficiently
neutralized ALV (SVVU-I 01) isolated in this study
Thc nucleotide sequence of gp85 and gp37 was determined for tht: field
isolate SVVU- 10 I and compared with publishcd sequences of' ALV subgroups A. B.
C. D, E and J and seven strains of' ALV subgroup-.I. The result of prrsent study
showed that the SVVU- I0 I belongs to ALV subgroup-J.
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