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/Usporedna procjena razlicitih postupaka lancane reakcije polimerazom uz prethodnu reverznu transkripciju za dokaz gena Fi N virusa kuge malih prezivaca u klinickim uzorcima

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 specificity 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--ELIISA 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. Key words: evaluation, molecular detection, peste des petits ruminants, reverse transcription polymerase chain reaction Istrazena je dijagnosticka vrijednost razlicitih pocetnica za gene F i N koje se sada rabe za dijagnosticiranje kuge malih prezivaca lancanom reakcijom polimerazom uz prethodnu reverznu transkripciju u usporedbi sa sendvic imunoenzimnim testom (S-ELISA). Testirano je ukupno pet parova pocetnica: dva para (F1/F2 i Fb1/Fb2) specificna za razlicita podrucja gena F i tri para specificna za razlicita podrucja gena N (NP3/NP4, pprn_fr2/pprn_rev i N1/N2). Njihova vrijednost bila je usporedena pretragom 10 klinickih uzoraka (cetiri uzorka krvi, cetiri uzorka nosnog obriska i dva uzorka tkiva) uzetih od zivotinja pod sumnjom na kugu malih prezivaca. Uporabom pocetnica NP3/NP4 dokazan je najveci broj pozitivnih uzoraka (6/10), a potom pocetnicom N1/N2 (5/10). Objema pocetnicama za gen F (F1/F2 i Fb1/Fb2) dokazana su