[ABSTRAK
Aeromonas hydrophila merupakan foodborne dan waterborne patogen, yang banyakditemukan pada lingkungan perairan. Bakteri ini dapat menginfeksi manusia, hewanteresterial dan hewan air seperti ikan mas, Infeksi A. hydrophila dapat mengakibatkankomplikasi gastrointestinal dan non-gastrointestinal pada manusia dengan penularanterjadi secara oral maupun luka yang terinfeksi bakteri. Sedangkan infeksi A.hydrophila pada ikan mas dapat menjadi sumber penularan ke manusia danmengakibatkan kematian ikan mas budidaya yang berdampak pada kerugian ekonomi.Hingga saat ini informasi mengenai infeksi A. hydrophila pada manusia masih jarangdilaporkan. Hal tersebut dapat terjadi karena hingga saat ini metode diagnosa antibodianti A. hydrophila belum banyak berkembang, sedangkan uji kekebalan penting bagiskrining, diagnosa banding dan uji konfirmasi terhadap infeksi A. hydrophila. Untukitu penelitian ini bertujuan untuk mengembangkan metode ELISA dan uji deteksicepat berdasarkan modifikasi ELISA menggunakan imunostik untuk mendeteksiantibodi anti A. hydrophila. Sebagai hewan uji digunakan enam ekor kelinci (NewZeland White) dan enam ekor ikan mas (Cyprinid carpio) yang diimunisasi denganantigen sel utuh bakteri A. hydrophila yang diiaktivasi dengan formalin 0,3%.Imunisasi pada ikan mas dilakukan secara intraperitoneal, diulang satu minggu setelahimunisasi pertama, sedangkan imunisasi pada kelinci dilakukan satu kali denganemulsi antigen dan Freund?s complete adjuvant kemudian dilanjutkan dengan dua kalibooster menggunakan emulsi antigen dan Freund?s incomplete adjuvant. Koleksiserum hewan uji dilakukan setiap minggu hingga minggu ke-6 dari koleksi serumpertama. Hasil optimasi terhadap uji ELISA menunjukkan bahwa uji ELISA yangdikembangkan mampu mendeteksi antibodi anti A. hyrophila, demikian pula denganimunostik yang dirakit mampu memdeteksi antibodi anti A. hyrophila pada serum ikanmas dan kelinci.
ABSTRACT
Aeromonas hydrophila is foodborne and waterborne pathogens. The bacteria thatoccur ubiquitously and autochthonously in aquatic environments. These bacteria caninfect humans, animals terrestrial and aquatic animals such as goldfish, infection of A.hydrophila can lead to complications of gastrointestinal and non-gastrointestinalinfection in humans and transmitted by oral and infected wounds contamination. Theinfected carp with A. hydrophila is a source of transmission to humans, resulting in thedeath of farmed fish that have an impact on economic disadvantage. Till now there arelacking information of A. hydrophila infection in humans reported. This can be occurbecause to the current diagnostic methods antibodi A. hydrophila undeveloped, whilethe immunity test is important for screening, differential diagnosis and confirmationtest of A. hydrophila infection. Therefore, the aims of this research is to developELISA methods and rapid detection test based on the modified ELISA usingimunostick to detect antibodies against A. hydrophila. For animals models, 6 rabbits(New Zeland White) and 6 carp (Cyprinid carpio) were immunized with whole cellantigen A. hydrophila bacteria which inactivated using 0.3% formalidehide. Carpimmunized intraperitoneally with antigen, and repeated one week after the firstimmunization, whereas immunization in rabbits done once with antigen emulsion andFreund's complete adjuvant followed by two booster using emulsion of antigen andFreund's incomplete adjuvant. Collection of animal serum done every week, till thesixth week from the first serum collection. The results of the optimization of theELISA test showed that the developed ELISA test is able to detect antibodies againstA. hyrophila, as well as the assembled imunostik capable to detect antibodies againstA. hyrophila both in carp and rabbit serum respectively, Aeromonas hydrophila is foodborne and waterborne pathogens. The bacteria thatoccur ubiquitously and autochthonously in aquatic environments. These bacteria caninfect humans, animals terrestrial and aquatic animals such as goldfish, infection of A.hydrophila can lead to complications of gastrointestinal and non-gastrointestinalinfection in humans and transmitted by oral and infected wounds contamination. Theinfected carp with A. hydrophila is a source of transmission to humans, resulting in thedeath of farmed fish that have an impact on economic disadvantage. Till now there arelacking information of A. hydrophila infection in humans reported. This can be occurbecause to the current diagnostic methods antibodi A. hydrophila undeveloped, whilethe immunity test is important for screening, differential diagnosis and confirmationtest of A. hydrophila infection. Therefore, the aims of this research is to developELISA methods and rapid detection test based on the modified ELISA usingimunostick to detect antibodies against A. hydrophila. For animals models, 6 rabbits(New Zeland White) and 6 carp (Cyprinid carpio) were immunized with whole cellantigen A. hydrophila bacteria which inactivated using 0.3% formalidehide. Carpimmunized intraperitoneally with antigen, and repeated one week after the firstimmunization, whereas immunization in rabbits done once with antigen emulsion andFreund's complete adjuvant followed by two booster using emulsion of antigen andFreund's incomplete adjuvant. Collection of animal serum done every week, till thesixth week from the first serum collection. The results of the optimization of theELISA test showed that the developed ELISA test is able to detect antibodies againstA. hyrophila, as well as the assembled imunostik capable to detect antibodies againstA. hyrophila both in carp and rabbit serum respectively] |
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