PCR [polymerase chain reaction]-based detection system for the diagnosis of Haemophilus paragallinarum infection
2003
Molina, H. A. | Torres, M. I. P. | Magbanua, J. P. V. | Rondina, M. M. R. | Balagat, A. M. | Alip, T. G., Philippines Univ. Los Banos, College, Laguna (Philippines). National Inst. of Molecular Biology and Biotechnology
Collection and isolation of HP8 [Haemophilus paragallinarum] from suspected IC [infectious coryza] cases was started Nov 2001, a total of 661 samples were collected from different places in Luzon (Batangas, Quezon, Palawan, Laguna and Ilocos Sur), Visayas (Samar and Cebu) and Mindanao (Surigao del Sur) [Philippines]: 389 from poultry layers, 11 from poultry breeders, 213 from gene fowl, 22 from Kabir and 26 from native chickens. So far, fifteen H. paragallinarum were isolated, 7 of which were obtained from poultry layers, 5 from game fowls and 2 from native chicken (Feb-Oct 2002). Biochemical characterization of these isolates has been done. Antisera production of serotypes A, B and C in rabbits induced high liters of 1:2048 for HPg serotypes A and B and 1:256 for HPg serotype C (Jan to March 2003). The procedure for DNA isolation of HPg using DNAzol (Gibco, Life Science) has been found to produce insufficient yield, even with the use of 8 mM NaOH to dissolve the DNA pellet. The method recommended by Dr. Blackall has been found to produce more satisfactory yield (Oct 2003). Trials using this method confirmed that Dr. Blackall's recommnded method for DNA isolation gave satisfactory yield (Jan 2003). The PCR protocol HPGR-PCR failed to detect the organism from colony preparation and extracted DNA, despite numerous modifications done. It has however, detected the organism from nasal and palatine cleft swab samples of chicken inoculated with the same organism. Repeated trials confirmed earlier results that HPg organism from nasal and palatine cleft swab samples of experimentally infected chickens were detected by HPGR-PCR. HPg in virulence state are detected by HPGR-PCR (Jan-March 2003).
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