‘ig. 1. (A) Result of the LAMP assay subjected to different temperatures. Lane 1, DNA marker; lane 2, negative control; and lanes 3-7, result of LAMP assay at different incubation emperature (in °C). (B) LAMP assay subjected at different incubation time. Lane 1, DNA marker; Lane 2, negative control; lanes 3-5, LAMP assay at different incubation time in minutes). The CAE, a disease in goats, has caused a damage that has a big impact in the goat industry. Animals affected shows decrease in productivity. At the moment, there is no vaccine developed for this particular disease, and the only way to control the spread of the virus is through culling and sacrificing the animal (CFSPH, 2007). The CAEV causes a persistent disease in goats, which is characterized by encephalomalacia, pneumonia, arthritis, and mastitis. Encephaloma- lacia is usually seen among kids, while arthritis is more common in As shown in Fig. 1A, the LAMP assay for CAEV could be performed at any temperatures from 61 °C to 65 °C, and the result of the assay Fig. 2. (A) Result of the LAMP assay on gel electrophoresis and visual inspection by (B) addition of SYBR green dye. Five samples were positive for CAEV. (C) Result of the nested-PCR assay. Four samples were positive for CAEV (as indicated by arrows) using the 2 sets of primers. Saclay AM hire ma INOS oR Sa a a as aa Sp ee tess Acme casts Shuai caine as Ba teeta ai iiiiaemaiamen eo Eixanickat cata iieat Momma Since the introduction of LAMP assay in 2000, this technique became widely used in different fields including detection of different diseases both virus and bacteria such as PED virus (Ren and Li, 2011), Japanese encephalitis virus (Parida et al., 2006), Mycobacterium spp. The LAMP reaction can be performed by incubating the reagents, specific primers, and sample in a single tube. Simple equipment such as water bath or a heat block can be used for this molecular technique for the detection of diseases in particular. The property of the assay wherein the results can be analyzed by the naked eye makes it a good diagnostic tool that could be used outside the laboratory or field. Result of the detection of CAEV by LAMP and nested-PCR assays. Table 2 adults. This virus in goats is believed to be transmitted directly by ingestion of colostrum and milk contaminated with the virus, while direct contact with the infected animal also considered a way of acquiring the disease. In a study by Villoria et al. (2013), CAEV was detected in air and water samples collected from the pen of infected animals. A PCR assay was used as a detection tool in the mentioned study. Fig. 3. Evaluation of specificity. (A) Visual inspection with SYBR green dye. Samples 1-3: DDW, PED, and FMD samples, while 4 is the CAEV sample. (B) Gel electrophoresis result, M - DNA ladder. Only sample 4 appeared positive after LAMP assay as indicated by the color change (A) and multiple ladder-like band (B).
