Bushmeat Species Identification: Recombinase Polymerase Amplification (RPA) Combined with Lateral Flow (LF) Strip for Identification of Formosan Reeves' Muntjac (Muntiacus reevesi micrurus).

The identification of animal species of meat in meat products is of great concern for various reasons, such as public health, religious beliefs, food allergies, legal perspectives, and bushmeat control. In this study, we developed a new technique to identify Formosan Reeves' muntjac in meat using recombinase polymerase amplification (RPA) in combination with a lateral flow (LF) strip. The DNA extracted from a piece of Formosan Reeves' muntjac meat was amplified by a pair of specific primers based on its mitochondrial cytochrome b gene for 10 min at a constant temperature ranging from 30 to 45 °C using RPA. Using the specific probe added to the RPA reaction system, the amplified products were visualized on the LF strip within 5 min. The total operating time from quick DNA extraction to visualizing the result was approximately 30 min. The RPA-LF system we designed was efficient when using boiled, pan-fried, roasted, stir-fried, or stewed samples. The advantages of simple operation, speediness, and cost-effectiveness make our RPA-LF method a promising molecular detection tool for meat species identification of either raw or variously cooked Formosan Reeves' muntjac meat. It is also possible to apply this method to identify the meat of other wildlife sources.

Development and validation of a new loop-mediated isothermal amplification for detection of pathogenic Leptospira species in clinical materials.

Leptospirosis, a global zoonotic disease, is often neglected but has a significant impact on human health. Here we reported a new loop mediated isothermal amplification (LAMP) assay targeting lipL32 gene of pathogenic Leptospira spp. Polymerase chain reaction (PCR), nested-PCR and real-time PCR assays were included in this study for the comparison of analytic and diagnostic sensitivity and specificity. LipL32 LAMP we designed enables detection of 10 copies of L. interrogans and has a higher diagnostic sensitivity (91.67%) and specificity (100%) than other PCR-based methods. The high sensitivity, specificity and flexible reaction conditions of the lipL32 LAMP assay makes it feasible for resource-limited countries and on-site application.

High prevalence of caprine arthritis encephalitis virus (CAEV) in Taiwan revealed by large-scale serological survey.

In this study, a large-scale serological survey of caprine arthritis encephalitis virus (CAEV) infection was conducted between March 2011 and October 2012. 3,437 goat blood or milk samples were collected from 65 goat farms throughout Taiwan. A commercial ELISA kit was used to detect antibodies against CAEV. The overall seropositive rate was 61.7% (2,120/3,437) in goats and in 98.5% (64/65) of goat farms. These results provide the first large-scale serological evidence for the presence of CAEV infection, indicating that the disease is widespread in Taiwan.

Serological and PCR detection of feline leptospira in southern Taiwan.

Taiwan is in the subtropical zone and has typhoons every year. Leptospirosis is an endemic disease in Taiwan, and feline leptospirosis in Taiwan remains unknown so far. From January, 2010, to September, 2011, 233 cats in south Taiwan (159 stray cats and 74 household cats) were sampled in this research. Leptospira antibody titer was detected by the serology gold standard, the microscopic agglutination test (MAT). Both serum and urine were examined for Leptospira DNA by polymerase chain reaction (PCR) with two sets of primers. In this study, the serological survey showed 21 (9.3%) examined sera contained antibodies specific for pathogenic Leptospira serogroups. The results of PCR revealed that 25 (19.1%) serum and 80 (67.8%) urine samples were found positive for leptospiral DNA sequences. All products amplified from PCR reactions were sequenced by an automated method for further confirmation. This is the first study concerning the epidemiology of pathogenic Leptospira in stray and household cats' urine, and the results demonstrate that some of the cats are susceptible to pathogenic Leptospira and have the potential to shed pathogenic Leptospira into the environment. This could be an issue of public health.