Avian influenza virus is a poultry viral disease, which causes high economic losses. Various efforts have been made to control the disease. One effort required fast and accurate screening diagnostic test. This study aimed to determine the potential of a rapid test kit, namely AIV/H5 Anigen Rapid Test for the detection of AI virus types A and subtype H5 in the fields. Some tests were carried out, e.g. the potential test, crossreaction test, sensitivity and specificity test. The potential test was done to evaluate detection limits of the kit, by having the test of a serial dilution of AI virus positive control. Crossreaction test was done to detect antigens other than AI virus H5N1, e.g. IB virus of Massachusetts strain, IBV strain 491, Newcastle Disease virus, and Escherichia coli. Sensitivity and specificity test were applied to the field samples which clinically and laboratory were confirmed as H5N1 positive. To confirm the result of rapid test was then being done by reverse transcriptase polymerase chain reaction. Based on these results it can be concluded that Anigen Kit AIV/H5 Ag Rapid Test can detect antigencontaining samples having AI virus HA titer up to 2 6 of type A virus, and up to 2 5 for subtype H5 virus. Anigen Kit AIV/H5 Ag Rapid Test showed no crossreactions with Infectious Bronchitis virus, Newcastle Disease virus, and Escherichia coli. Anigen A Rapid Test Kit AIV Ag showed a sensitivity of 50% and specificity of 100%, while Anigen Ag Rapid Test Kit AIV/H5 showed a sensitivity of 25% and specificity of 100%.
Red sea bream iridoviral disease (RSIVD) infection is known as a contagious disease in the marine aquaculture commodities mainly on grouper (Epinephelussp.)which causes a high mortality rate. Symptoms of disease were weak, darker skin and swollen spleen of fish. The aim of this study was to create and apply a rapid diagnostic test supported by molecular analysis. Field trials on a mass mortality outbreaks were identified in the city of Tanjungpinang, Indonesia. Serum anti RSIV was obtained by immunizing of the vaccine RSIV intraperitoneally on rabbits with graded doses per week was 0.5, 1, 2 and3mL, to boost antibody titers. In the fifth week, serum was harvested via the auricular vein; serum was purified to obtain immunoglobulin G(IgG)then was coupling with protein A ofStaphylococcus aureus at the same volume(kit co-agglutination RSIVD). Field samples of spleen were taken from the normal fish and suspected fish then crushed and suspended with PBS pH 7.2, and centrifuged at 8.000 rpm for 15 min. Fifty microliters of RSIVDco-agglutination kit and 50 µL of spleen supernatant were reacted on the sterile glass object. The results showed sandy agglutination after 10 min for positively infected spleen, and no agglutination in the samples of healthy fish (negative) as well as in control with PBS (negative). Confirmation testing by polymerase chain reaction (PCR) using primer forward 1-F (5’-CTC-AAA-CAC-TCT-GGC-TCA-TC-3’) and reverse 1-R (5’-GCA-CCA-ACA-CAT-CTC-CTA-TC-3’) had a band of 570 bp. Sequencing results showed the similarity of 99% identity with RSIV. Testing with RSIVD co-agglutination kit showed the advantages such as cheap, fast and an accurate in diagnosing the red bream iridoviral disease (RSIVD).
Penelitian ini bertujuan mengetahui potensi kit deteksi cepat Anigen® rapid test kit rabies Ag dalam mendeteksi virus rabies pada sampel otakanjing yang diperoleh dari lapangan yang meliputi batas deteksi, kecepatan reaksi, uji reaksi silang, uji sensitivitas, dan spesifisitas. Batas deteksi ditentukan dengan pengenceran secara serial kontrol positif virus rabies dan selanjutnya diuji dengan rapid test kit sesuai petunjuk produsen. Uji reaksi silang dilakukan dengan canine parvovirus, Escherichia coli, dan Salmonella sp. Uji sensitivitas dan spesifisitas dilakukan terhadap sampel otak yang telah dikonfirmasi positif rabies dengan uji fluorescent antibody technique. Konfirmasi uji rapid test tersebut dilakukan dengan reverse transcriptase polymerase chain reaction. Berdasarkan data yang diperoleh dalam penelitian ini dapat disimpulkan bahwa Anigen® rapid test kit rabies Ag mampu mendeteksi sampel yang mengandung virus rabies dengan titer 0,5 x log 106,5/0,03 ml, dengan rata-rata kecepatan reaksi 1,8 menit 29,35 detik (kurang dari 2 menit). Di samping itu Anigen® rapid test kit rabies menunjukkan tidak terdapat reaksi silang dengan canine parvovirus, Escherichia coli, dan Salmonella sp. serta mempunyai sensitivitas 92,30% dan spesifisitas 97,90%
Injure healing process occured on four phases such as hemostatis, inflammation, and maturation. Latex of Patah Tulang (Euphorbia tirucalli L.) can heal the wound because of its component active. Aim of this research was to find out the effect of ointment topical of E. tirucalli L. to wound healing process based on macroscopic and microscopic examination and cellular reaction of CD4+ lymphocyte. Male Wistar Rat, 2 month old, 150-200 Gram of body weight, as much as 36 rats. Rats were divided into 4 groups, and were anesthesized before punctioned on the 2 sides of lateral vertebrae. Group one was used as a control, group two was given with the ointment 9% of E. tirucalli L., group three was given with 23% of E. tirucalli L., and group four was given madecassol®. Treatment was given once a day for 15 days. Macroscopical and microscopical changes were observed at the fifth, tenth, and fifteenth day. Macroscopical observation was the diameter of injure. Microscopical observation consisted of epidermal thicness, number of fibroblast, using Haematoxylin and Eosin staining, and cellular reaction of CD4+ lymphocyte using imunohistochemistry. The results showed that topical administration of E. tirucalli ointment 9% and 23% potential to accelerate wistar skin wound healing puncture wounds. There was no difference between E. tirucalli ointment 9% and 23% application to observed parameters.Keywords: Euphorbia tirucalli L., injure healing, macroscopic, microscopic, lymphocyte CD4+
Kasus penyakit Infectious Bursal Disease (IBD) dewasa ini masih sering ditemukan pada peternakan ayam komersial baik layer maupun broiler di Indonesia. Diagnosis penyakit IBD sejauh ini mengandalkan lesi patologik spesifik dan kultur in ovo dengan mengamati lesi makroskopis embrio, serta diidentifikasi dengan uji agar gel presipitasi (AGP). Penelitian ini bertujuan menerapkan diagnosis dengan teknik reverse transcriptase polymerase chain reaction (RT-PCR) dari sampel Bursa Fabrisius (BF) sebagai konfirmasi pada kasus terdiagnosa IBD. Deteksi serologis virus IBD dengan uji AGP dengan sumber antigen chorioallantoic membrane (CAM) dan embrionya, untuk melihat potensinya sebagai sumber antigen uji AGP. Sampel BursaFabrisius sebanyak 5 yang diperoleh pada kasus terdiagnosa IBD, dikoleksi dari peternakan ayam komersial di Yogyakarta. Konfirmasi diagnosis dilakukan dengan metode RT-PCR. Sampel positip uji RT-PCR yang mengamplifikasi fragmen gen VP2. Isolasi virus IBD yang dilakukan kultur in ovo pada telur ayam berembrio (TAB) antibodi negatif terhadap virus IBD, berumur 11 hari. Desposisi materi inokulasi dilakukan pada (CAM), diinkubasi selama lima hari. Panen virus dilakukan dengan mengkoleksi membran korioalantois dan embrio, selanjutnya diamati lesi makroskopis yang timbul akibat infeksi virus IBD. Membran korioalantois dan embrioselanjutnya digerus dan diproses sebagai suspensi antigen yang digunakan dalam uji AGP. Hasil uji RT-PCR terhadap lima sampel Bursa Fabrisius yang dikoleksi dari peternakan ayam terdiagnosa penyakit IBD, tiga sampel menunjukkan hasil positif teramplifikasi fragmen gen VP-2 virus IBD dengan produk amplifikasi sebesar 440 bp, sedangkan dua sampel sisanya menunjukkan hasil negatif. Uji AGP dengan sumber antigen CAM menunjukkan hasil positip 2 dari 3 sampel yang diuji, sedangkan sumber antigen embrio menunjukkanhasil negatif. Berdasarkan data yang diperoleh dalam penelitian ini dapat disimpulkan bahwa uji RT-PCR dapat digunakan dalam mendeteksi virus IBD dari sampel BF terdiagnosa IBD. Uji AGP dengan sumber antigen CAM menunjukkan hasil lebih baik dari pada embrionya.
Abstract:Iridovirus infection often causes death and considerable economic losses in the aquaculture industry. This research applies the co-agglutination method that is fast, cheap and accurate in confirming the diagnosis of the cause of an outbreak of illness caused by iridovirus in the field, so that remedial action can be taken quickly and appropriately to minimize the impact of wider losses. Samples were taken from the grouper and pomfret star farms that are experiencing outbreaks of infectious diseases in the months from May to August 2015, Tanjungpinang, Indonesia. The sick and allegedly attacked by iridovirus samples showed abnormal swimming clinical symptoms, weakness and the swollen spleen. The swollen spleen of sick fish created suspension in phosphate buffered saline (PBS) with pH 7.2, and then centrifuged at 8,000 rpm for 10 min. The supernatant after centrifuge was used as the test sample. On a clean object glass, 50 µL of the supernatant was treated with 50 µL kit co-agglutination pre-prepared. The positive results were shown by the agglutination reaction after 10-15 min, while as a negative control, PBS was reacted with co-agglutination kit that looked homogeneous (no agglutination). It was showed that the grouper (Epinephelus sp.) on four farms and pomfret star (Thracinotus blochii) on one farm that experienced an outbreak of infectious disease in Tanjungpinang showed positively infected iridovirus. The same positive iridovirus result was also demonstrated by examination using polymerase chain reaction (PCR) at 570 bp. So, the causative agent of plague on grouper and pomfret star was iridovirus. In addition, the co-agglutination test based on serology is more quick, cheap and accurate.
For the past few years, Edwardsiella tarda has become major problem in African catfish culture in Jambi. Detection by biochemical characteristic can lead to inaccurate result and there is a necessity to develop more specific and accurate method, one of which is Agar Gel Precipitation (AGP) method. Six samples each were collected from two African catfish farm located in District Sungai Gelam and Telanai Pura in Jambi, which was showed clinical signs of E. tarda outbreak with more than fifty percent mortality rate. Heat stable soluble antigen was prepared from 2 groups of pure culture isolated from sample for AGP test. Antiserum for test wells was antiserum of E. tarda (ATCC 15947) that have been produced by inoculating whole-cell antigen (heat-stable) and flagellar antigen (formalin-killed) in rabbit. For control positive, soluble antigen prepared from E. tarda (ATCC 15947), and control negative from Aeromonas hydophila (ATCC 35654) and Edwardsiella ictaluri (NCIMB 13272). Both antiserums were able to show positive reaction visible by the formation of specific precipitin lines between antiserum and antigen wells, and there was no precipitin reaction for negative control. In conclusion AGP method is a one of reliable technique to identify E. tarda.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.