• Journal of Veterinary Science
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• v.21 no.2
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• pp.14.1-14.10
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• 2020

African swine fever (ASF) is a highly contagious disease of domestic pigs and wild boars (WBs). Without a vaccine, early antibody and antigen detection and rapid diagnosis are crucial for the effective prevention of the disease and the employment of control measures. In Sardinia, where 3 different suid populations coexisted closely for a long time, the disease persists since 1978. The recent ASF eradication plan involves more stringent measures to combat free-ranging pigs and any kind of illegality in the pig industry. However, critical issues such as the low level of hunter cooperation with veterinary services and the time required for ASF detection in the WBs killed during the hunting season still remain. Considering the need to deliver true ASF negative carcasses as early as possible, this study focuses on the evaluation and validation of a duplex pen-side test that simultaneously detects antibodies and antigens specific to ASF virus, to improve molecular diagnosis under field conditions. The main goal was to establish the specificity of the two pen-side tests performed simultaneously and to determine their ability to detect the true ASF negative carcasses among the hunted WBs. Blood and organ samples of the WBs hunted during the 2018/2019 hunting seasons were obtained. A total of 160 animals were tested using the pen-side kit test; samples were collected for virological and serological analyses. A specificity of 98% was observed considering the official laboratory tests as gold standards. The new diagnostic techniques could facilitate faster and cost-effective control of the disease.

• Journal of Bio-Environment Control
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• v.26 no.4
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• pp.333-339
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• 2017

This research was aimed to establish rapid analysis technique for the determination of nitrate ($NO_3{^-}$) concentration in the leaves of paprika, which has key role for the stable vegetative and reproductive growth. Leaf petiole and blade sap of two paprika cultivars ('Raon red' and 'Raon yellow') were used for the determination of $NO_3{^-}$ concentration, separately using rapid detection kit (RQ-flex) and spectroscopy quantification methods. In addition, two paprika cultivars namely, 'Nicole' and 'TP2001' were used to determine the status of $NO_3{^-}$ concentration in leaf of each fruiting group. $NO_3{^-}$ concentration in leaf blade sap and the content in leaf showed significant correlation ($R^2=0.8628$), analysed by RQ-flex and spectroscopy methods, respectively. Furthermore, leaf petiole sap and the content in leaf also showed significant correlation ($R^2=0.6734$) but the relationship was poor compared to leaf blade sap and the leaf content. $NO_3{^-}$ concentration in petiole sap decreased in all the cultivars from early to late fruiting group. The higher concentration in the lower leaves and the continuous decrease towards the upper leaves in the both years were found through the analysis of $NO_3{^-}$ concentration in different leaf position. In addition, daily short-term fluctuation of $NO_3{^-}$ in petiole sap could be rapidly monitored. These results showed that long-term or short-term monitoring by test strip-based rapid analysis technique might be useful tool for the diagnosis of nutritional status for the stable of nutritional management in paprika.

• Korean Journal of Veterinary Service
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• v.35 no.4
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• pp.333-337
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• 2012

This study was carried out to investigate the prevalence of bovine leukemia virus (BLV) infection and to compare the results of enzyme-linked immunosorbent assay (ELISA) and nested polymerase chain reaction (nPCR) in dairy farms in northern Gyeonggi province from August through December 2011. A total of 625 dairy cattle from 14 dairy farms were tested for antibodies against BLV using commercially available ELISA test kit. The overall seroprevalence of BLV infection was 76.3%. The seroprevalence of diary cattle according to age was the highest at 61~72 months (88.0%, P<0.001). Two hundred fifty one dairy cattle from 7 diary farms were tested ELISA and nPCR. The kappa value of BLV between ELISA and nPCR was 0.765. The results indicate that BLV infection spread widely in dairy farms and the nPCR is rapid method for the early detection of BLV infection.

• Biomedical Science Letters
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• v.15 no.4
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• pp.295-300
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• 2009

Tuberculosis caused by Mycobacterium tuberculosis (MTB) still remains to be the most dreadful infectious disease affecting almost every country. In the present study, we developed a simple and rapid but accurate and sensitive assay method for detecting MTB using microplate hybridization assay. For this, a selective region of the rpoB gene was used to design PCR primers, and MTB and Mycobacterium genus-specific probe molecules. The specificity of the assay was confirmed using fifteen different mycobacterial reference strains and twelve different non-mycobacterial reference strains, and the sensitivity was determined to be 100 fg using genomic DNA of MTB reference strain, H37Rv. Subsequently, a total of 62 sputum samples with diverse smear scores and culture positive results were used to evaluate the kit performance. In brief, the specificity and the sensitivity of the assay were 100% and 98.4%, respectively.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1350-1358
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• 2012

Viral pathogens, alongside other pathogens, have major effects on crustacean aquaculture. Hepatopancreatic parvovirus (HPV) is an emerging virus in the shrimp industry and has been detected in shrimp farms worldwide. The HPV genome has greater diversity than other shrimp viruses owing to its wide host range and geographical distribution. Therefore, developing diagnostic tools is essential to detect even small copy numbers from the target region of native HPV isolates. We have developed two easy to use quantitative real-time PCR kits, called Green Star and Dual Star, which contain all of the necessary components for real-time PCR, including HPV primers, using the primers obtained from the sequences of HPV isolates from Korea, and analyzed their specificity, efficiency, and reproducibility. These two kits could detect from 1 to $1{\times}10^9$ copies of cloned HPV DNA. The minimum detection limits obtained from HPV-infected shrimp were $7.74{\times}10^1$ and $9.06{\times}10^1$ copies in the Green Star and Dual Star assay kits, respectively. These kits can be used for rapid, sensitive, and efficient screening for HPV isolates from Korea before the introduction of postlarval stages into culture ponds, thereby decreasing the incidence of early development of the disease.

• Korean Journal of Clinical Laboratory Science
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• v.41 no.4
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• pp.145-152
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• 2009

Novel influenza A virus, subtype H1N1 of swine-lineage, has been transmitted rapidly to many regions of the world. Rapid detection of the virus is essential to instigate appropriate patient care and public health management and for disease surveillance. The aim of this study is to determine the prevalence of novel influenza A (H1N1) virus in Korea using reverse-transcription real time polymerase chain reaction (rRT-PCR). Novel H1N1 virus was detected in a total of 8,948 nasopharyngeal samples from patients with influenza-like illness throughout Korea from August to September 2009. RNA was extracted from $300{\mu}l$of sample using an RNA extraction kit (Zymo Research, CA, USA). In the present study, Genekam kit (Genekam, Duisburg, Germany) was used to detect novel H1N1 virus. Novel H1N1 virus was found in 1,130 samples from a total of 8,948 samples (12.6%). The highest frequency was found in 10- to 19-year-olds (M: 29.3% vs. F: 16.4%), followed by 20- to 29-year-olds (M: 17.9% vs. F: 15.4%), 40- to 49-year-olds (M: 6.5% vs. F: 8.1%), 50- to 59-year-olds (M: 6.0% vs. F: 5.5%), and 30- to 39-year-olds (M: 4.6% vs. F: 3.8%). The mean positive rate was higher in men than in women (M: 14.7% vs. F: 7.4%). Novel H1N1 virus showed the lowest prevalence in patients over 60 years old. The positive rate increased daily and showed a significant high peak in mid-September 2009. In 19 provinces of Korea, Cheonan (41.1%), Busan (37.3%), Gangneung (33.3%), Jinju (32.1%), Ulsan (24.6%), Deajeon (23.7%) areas showed high frequencies and other provinces were found less than 10% of novel H1N1 virus. Since reverse-transcription real time PCR assay is rapid, accurate, and convenient, it may assist public health laboratories in detecting novel H1N1 virus. Moreover, these data could be useful for the management of patients with influenza-like illness.

• Parasites, Hosts and Diseases
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• v.52 no.2
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• pp.143-149
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• 2014

To evaluate the seroprevalence against circumsporozoite protein (CSP) of Plasmodium vivax in sera of Korean patients, the central repeating domain (CRD) of CSP was cloned and analyzed. From the genomic DNA of patient's blood, 2 kinds of CSPs were identified to belong to a VK210 type, which is the dominant repeating of GDRA(D/A)GQPA, and named as PvCSPA and PvCSPB. Recombinantly expressed his-tagged PvCSPA or PvCSPB in Escherichia coli reacted well against sera of patients in western blot, with the detecting rate of 47.9% (58/121), which included 15 cases positive for PvCSPA, 6 cases positive for PvCSPB, and 37 cases for both. The mixture of PvCSPA and PvCSPB was loaded to a rapid diagnostic test kit (RDT) and applied with the same set of patient sera, which resulted in detection rates of 57.0% (69/121). When the protein sequences of PvCSPA were compared with those of P. vivax in endemic regions of India and Uganda, they were compatibly homologous to PvCSPA with minor mutations. These results suggested that the recombinant PvCSPA and PvCSPB loaded RDT may be a milestone in latent diagnosis which has been a hot issue of domestic malaria and important for radical therapy in overlapped infections with P. falciparum in tropical and subtropical areas. During the biological process of malarial infection, exposure of CSP to antigen-antibody reaction up to 57.0% is the first report in Korea.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.12
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• pp.1746-1750
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• 2004

The PBM ${BioSign}^{TM}$ Salmonella (PBMS) test kit based on an mmunochromatographic method was evaluated for the screening of Salmonella spp. in pure cultures, and 80, 15, and 10 artificially and naturally contaminated, and negative controlled food samples, respectively. The PBMS test involves presumptive qualitative procedures, detecting the presence of Salmonella spp. in foods within 26 h total testing period and allowing the user to release negative products 70 h earlier than the conventional methods. The PBMS test using Buffered Peptone Water and Rappaport-Vassiliadis broth was evaluated for 10 different food types for various Salmonella spp. It showed detection limits of 1 to 25 colony forming units (CFU)/25 g. No cross-reaction was observed, particularly to other gramnegative bacteria. These results indicate the PBMS test is a rapid and inexpensive procedure for the screening of Salmonella spp. present at low concentrations (1 to 25 CFU/25 g) in foods.

• Parasites, Hosts and Diseases
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• v.52 no.3
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• pp.305-310
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• 2014

Ascidian soft tunic syndrome (AsSTS) caused by Azumiobodo hoyamushi (A. hoyamushi) is a serious aquaculture problem that results in mass mortality of ascidians. Accordingly, the early and accurate detection of A. hoyamushi would contribute substantially to disease management and prevention of transmission. Recently, the loop-mediated isothermal amplification (LAMP) method was adopted for clinical diagnosis of a range of infectious diseases. Here, the authors describe a rapid and efficient LAMP-based method targeting the 18S rDNA gene for detection of A. hoyamushi using ascidian DNA for the diagnosis of AsSTS. A. hoyamushi LAMP assay amplified the DNA of 0.01 parasites per reaction and detected A. hoyamushi in 10 ng of ascidian DNA. To validate A. hoyamushi 18S rDNA LAMP assays, AsSTS-suspected and non-diseased ascidians were examined by microscopy, PCR, and by using the LAMP assay. When PCR was used as a gold standard, the LAMP assay showed good agreement in terms of sensitivity, positive predictive value (PPV), and negative predictive value (NPV). In the present study, a LAMP assay based on directly heat-treated samples was found to be as efficient as DNA extraction using a commercial kit for detecting A. hoyamushi. Taken together, this study shows the devised A. hoyamushi LAMP assay could be used to diagnose AsSTS in a straightforward, sensitive, and specific manner, that it could be used for forecasting, surveillance, and quarantine of AsSTS.

• Journal of Microbiology and Biotechnology
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• v.17 no.7
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• pp.1152-1161
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• 2007

An immunochromatography (ICG) strip test based on a monoclonal antibody for the rapid detection of L. monocytogenes in meat and processed-meat samples was developed in this study. A monoclonal antibody (MAb) specific to L. monocytogenes was produced from cloned hybridoma cells (FKLM-3B12-37) and used to develop an ICG strip test. The antibody showed a stronger binding to L. monocytogenes than other Listeria species, and a weak cross-reaction to S. aureus based on an ELISA. The detection limit of the ICG strip test was $10^5\;cell/ml$. In total, 116 meat and processed-meat samples were collected and analyzed using both the ICG strip test and a PCR. The ICG strip test and PCR indicated L. monocytogenes contamination in 34 and 27 meat samples, respectively. The 7 meat samples not identified as L. monocytogenes positive by the PCR were also tested using an API kit and found to be contaminated by Listeria species. In conclusion, the ICG strip test results agreed well with those obtained using the PCR and API kit. Thus, the developed ICG has potential use as a primary screening tool for L. monocytogenes in various foods and agricultural products, generating results within 20 min without complicated steps.