The use of filter paper as a transport device for serology of Pasteurella multocida infection: Analysis and comparison of protein composition of filter paper extract and serum

英语. Two methods for collecting blood specimens for measuring antibody to Pasteurella multocida were compared. Blood was collected on filter-paper strips, air-dried and stored at 4 deg. C along with paired samples collected by venepuncture. Analysis using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the protein composition of filter paper extract and serum was similar. Both samples had common proteins of 67, 52-58 and 27 kDa. However, there are two proteins bands of 14 and 30 kDa that were only found in filter-paper extract. Westernblot analysis also showed that samples from both sampling techniques reacted to P. multocida proteins of 43 kDa. Samples from experimental and field animals were also collected by the two techniques and assayed by enzyme-linked immunosorbent assay (ELISA) for P. multocida antibodies. The agreement between samples from experimental animals and the field using ELISA was analyzed. Samples from experimental animals, showed a very high correlation (r = 0.931) in ELISA results among samples collected by the two techniques. However, the correlation was lower (r = 0.799) in samples collected from the field. Cost analysis showed that filter-paper collection technique was 100 times more economical compared to venepuncture technique. It was concluded that extracts of whole blood dried on filter paper can be used as an alternative to sera in ELISA for measuring antibodies to P. multocida

印度尼西亚. Dua cara pengambilan sampel darah untuk uji terhadap antibodi Pasteurella multocida dibandingkan dalam penelitian ini. Darah dikumpulkan dengan menggunakan kertas saring dan tabung. Komposisi protein ekstrak kertas saring dan serum dianalisis dengan sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Analisis ini menunjukkan bahwa protein dengan bobot molekul 67, 52-58 dan 27 kDa adalah komponen yang ditemukan pada kedua teknik pengambilan sampel. Tetapi, ada 2 komponen protein 14 dan 30 kDa yang hanya ditemukan pada ekstrak kertas saring. Pemeriksaan Westernblot membuktikan bahwa ekstrak kertas saring dan serum bereaksi sama terhadap P. multocida, yaitu pada protein 43 kDa. Pengamatan pada uji enzyme-linked immunosorbent assay (ELISA) menunjukkan terdapat hubungan yang erat antara tingkat densitas optikal sampel kertas saring dan serum dari hewan percobaan dan hewan lapangan. Pada sampel hewan percobaan, hubungan kedua teknik pengambilan sampel tersebut tinggi (r=0,931), sedangkan pada hewan lapangan hubungan tersebut lebih rendah (r=0,799). Perhitungan ekonomis terhadap biaya operasional pengambilan sampel di atas menunjukkan bahwa teknik kertas saring 100 kali lebih murah daripada sistem tabung. Disimpulkan bahwa ekstrak darah pada kertas saring dapat digunakan sebagai pengganti serum pada uji ELISA terhadap P. multocida