Izvorni znanstveni članak

Comparative evaluation of different F and N gene based reverse transcription polymerase chain reaction for molecular detection of peste des petits ruminants virus from clinical samples.

Sumit Mahajan ; Division of Veterinary Epidemiology and Preventive Medicine, SKUAST-Jammu, J&K, India
Rajesh Agrawal ; Division of Veterinary Epidemiology and Preventive Medicine, SKUAST-Jammu, J&K, India
Mahesh Kumar ; Department of Epidemiology and Preventive Medicine, GBPUA&T, Pantnagar, Uttrakhand, India
Anand Mohan ; Department of Epidemiology and Preventive Medicine, GBPUA&T, Pantnagar, Uttrakhand, India
Nishi Pande ; Division of Animal Reproduction Gynecology and Obstetrics, SKUAST-Jammu, J&K, India

Sažetak

In the present study the diagnostic value of different sets of F and N gene based primers currently used for diagnosis of Peste des petits ruminants (PPR) by reverse transcription polymerase chain reaction (RT-PCR) were assessed by comparing it with Sandwich ELISA (S-ELISA). A total of five primer pairs, consisting of two pairs (F1/F2 and Fb1/Fb2) amplifying two different regions of F gene and three pairs amplifying different regions of N gene (NP3/NP4, pprn_fr2/pprn_rev and N1/N2) were compared on 10 clinical samples (4 blood, 4 nasal swabs and 2 tissue samples) collected from animals suspected for PPR. The primer sets NP3/NP4 detected highest number of positive samples 6 out of 10 followed by N1/N2 (5/10). Both F-gene based primers (F1/ F2 and Fb1/Fb2) detected 3 out of 10 samples as positive. Whereas the primer pair pprn_fr2/pprn_rev did not yield the desired amplicon in any of the samples tested. The maximum sensitivity and specififi city of 100% was observed by NP3 and NP4 primer based RT-PCR whereas, 0% sensitivity was recorded by pprn_fr2/pprn_rev which fail to detect any positive sample. The overall agreement of 100% with kappa value 1.00 was highest between S-ELISA and NP3 and NP4 primer based RT-PCR suggesting an almost perfect agreement, followed by N1/N2, having kappa value of 0.800, suggesting a substantial agreement. Results thus obtained in the present study, suggest that F-gene primers based RT-PCR can be easily replaced by highly sensitive and specific N-gene primers based RT-PCR for detection of PPR virus nucleic acid.

Ključne riječi

evaluation; molecular detection; peste des petits ruminants; reverse transcription polymerase chain reaction

Hrčak ID:

128427

URI

https://hrcak.srce.hr/128427

Datum izdavanja:

10.10.2014.

Podaci na drugim jezicima: hrvatski

Posjeta: 1.594 *