• Bulletin of the Korean Chemical Society
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• 제25권5호
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• pp.757-760
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• 2004

• 한국동물위생학회지
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• 제29권2호
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• pp.103-110
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• 2006

Ticks cause economic losses to the deer industry by decreasing the growth and production of the farmed animals. The mediation of ticks affects humans and animals by causing contagious disease both directly and indirectly. Blood from farmed deer from the areas near Jangsu branch was collected for screening of infectious protozoa and rickettsial disease. Seventy deer blood samples were collected from 30 different deer farms located in Jinan, Jangsu and Muju. This blood samples were used for blood slide smear examination and hematological analysis. DNA from these samples was extracted and was used for PCR analysis for detection of gene fragments of Theileria spp, Babesia spp, Anaplasma spp and Ehrlichia spp. In the blood slide smear examination and PCR analysis all samples did not show presence of protozoal and rickettsial diseases. Eight blood samples showed anemia, 1 sample showed iron deficiency and 7 samples showed regenerative anemia. Results for PCR analysis showed 2 samples were positive for T orientalis. All DNA samples were negative for Babesia spp, Anaplasma spp, and Ehrlichia spp.

• Parasites, Hosts and Diseases
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• 제51권5호
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• pp.573-577
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• 2013

A loop-mediated isothermal amplification (LAMP) assay allows rapid diagnosis of Toxoplasma gondii infection. In the present study, the LAMP assay was evaluated using blood from both naturally and experimentally infected pigs. The sensitivity of the LAMP assay was compared with that of Q-PCR. Both assays detected T. gondii in the blood of experimentally infected pigs, with 100% agreement. In infected blood samples, the parasite was detected as early as 2 days post-infection and reached a peak in 3-5 days. In 216 field serum samples, the detection rates of LAMP and Q-PCR assays were 6.9% and 7.8%, respectively. This result indicates that the sensitivity of the LAMP assay was slightly lower than that of the Q-PCR assay. However, the LAMP may be an attractive diagnostic method in conditions where sophisticated and expensive equipment is unavailable. This assay could be a powerful supplement to current diagnostic methods.

• Parasites, Hosts and Diseases
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• 제47권3호
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• pp.227-233
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• 2009

Local malaria transmission in the United Arab Emirates (UAE) came to an end in 1997. Nevertheless, UAE has been subjected to substantial importation of malaria cases from abroad, concerning both UAE nationals and immigrants from malarious countries with a total number of 2,119 cases in 2007. To evaluate a new DNA extraction technique using nested PCR, blood samples were collected from 132 individuals who presented to Infectious Diseases Department in Rashid Hospital, Dubai, and Central Department of Malaria Control with fever and persistent headache. Giemsa-stained blood films and ELISA test for malaria antibodies were carried out for detection of Plasmodium infection. Plasmodium infections were identified with the genus-specific primer set and species differentiation using nested PCR. A rapid procedure for diagnosis of malaria infections directly from dried blood spots using for the first time DNA extract from FTA Elute cards was evaluated in contrast to extraction techniques using FTA classic cards and rapid boiling technique. Our new simple technique for DNA extraction using FTA Elute cards was very sensitive giving a sensitivity of 100% compared to 94% using FTA classic cards and 62% in the rapid boiling technique. No complex preparation of blood samples was required prior to the amplification. The production cost of DNA isolation in our PCR assay was much less incomparable to that of other DNA extraction protocols. The nested PCR detected plasmodial infection and could differentiate P. falciparum from P. vivax, and also detected the mixed infection.

• 한국임상수의학회지
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• 제31권1호
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• pp.57-60
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• 2014

6개월령의 한국토종단모고양이가 발열, 빈호흡, 식욕부진, 체중감소로 내원하였다. 고양이는 10일 전 Fellocel$^{(R)}$ 백신을 접종받은 이후 증상을 보였다. 신체검사 결과 공막의 황달과 호흡곤란을 호소했다. 혈액혈구검사와 혈액화학검사에서 빈혈, 혈소판감소증, 백혈구증가증, 고빌리루빈혈증이 관찰되었다. 혈액도말 검사와 기타 검사를 통해 심한 용혈을 확인할 수 있었고 고양이 용혈성 빈혈을 감별진단 하기위해 Erhlichia, Hemobartonella, FeLV, FIV, Anaplasma PCR 검사를 의뢰하였다. PCR 검사 결과 Mycoplasma haemofelis 특이 208 bp의 양성밴드가 확인되었고, 다른 질병은 음성으로 진단되었다. 고양이는 4주 동안 Doxycycline을 사용하였고 급성기 1주 동안에는 prednisolone을 같이 사용하였다. 임상증상은 약물투여 4일째부터 호전되기 시작하였고 이후 PCR 재검을 통해 완치를 확인하였다. 이것은 M. haemofelis 감염 고양이의 진단을 확인하고 완치한 국내 첫 증례이다.

• 대한수의학회지
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• 제36권3호
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• pp.607-614
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• 1996

Several methods for rapid and accurate detection of VTe-producing E coli were established. These methods contain beta-glucuronidase-secretion test, beta-haemolysis-production test in blood agar, verocytotoxicity test, and PCR. All of the VTe-producing strains made beta-haemolysis on 5% sheep blood agar. VTe-producing strains secreted beta-glucuronidase whereas 0157:H7 strains producing VTI or VTII did not secrete that enzyme. Verocytotoxicity test was established for rapid diagnosis. VTe detection was rapider in Vero cell suspension than Vero cell monolayer. In PCR, there was a positive result only in VTe-producing E coli, not in VTI or VTII-producing E coli. In this experiment, 165 strains of E coli were islated from feces or intestinal contents of post-weaning piglets showing nervous sign or diarrhea. And 20 strains of E coli that produced VTe were selected by verocytotoxicity test and PCR. According to these experiments, there was a direct correlation between verocytotoxicity test and PCR. And verocytotoxicity test is recommended as a routine diagnostic method and PCR does as a accurate diagnostic method to detect VTe-producing E coli.

• 대한바이러스학회지
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• 제27권1호
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• pp.49-57
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• 1997

We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.

• Journal of Microbiology and Biotechnology
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• 제25권2호
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• pp.255-267
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• 2015

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne's disease, a chronic debilitating disease affecting ruminants worldwide. In the present study, we aimed to determine the major gene networks and pathways underlying the immune response to MAP infection using whole-blood cells, as well as provide the potential transcriptional markers for identifying the status of MAP infection. We analyzed the transcriptional profiles of whole-blood cells of cattle identified and grouped according to the presence of MAP-specific antibodies and the MAP shed by them. The grouping was based on the results obtained by ELISA and PCR analyses as follows: i) Test1 group: MAP-negative results obtained by ELISA and positive results obtained by PCR; ii) Test2 group: MAP-positive results obtained by ELISA and negative results obtained by PCR; iii) Test3 group: MAP-positive results obtained by ELISA and positive results obtained by PCR; iv) uninfected control: MAP-negative results obtained both by ELISA and PCR analysis. The results showed down-regulated production and metabolism of reactive oxygen species in the Test1 group, activation of pathways related to the host-defense response against MAP (LXR/RXR activation and complement system) in the Test2 and Test3 groups, and anti-inflammatory response (activation of IL-10 signaling pathway) only in the Test3 group. Our data indicate a balanced response that serves the immune-limiting mechanism while the host-defense responses are progressing.

• International Journal of Industrial Entomology and Biomaterials
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• 제11권2호
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• pp.135-138
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• 2005

Two pairs of DNA primers were designed for the detection of the Zhenjiang (China) strain of Bombyx mori densonucleosis virus (BmDNV-Z). These primers were designed from the nucleotide sequence of major structural protein gene (putative VD1-ORF2). PCR amplification was attempted from different issues (including silk gland, blood, skin and midgut) and feces of the silkworm which infected wit BmDNV-Z were amplified by PCR. Both of the primers gave expected size of in the DNA bands from midgut and feces, but not in the DNA of silk gland, blood and skin. The two bands were sequenced, and their sequence were same as the sequence designed for. BmDNV-Z could be successfully detected in single silkworm after it was infected for 12 hrs, and could not be detected before 9 hrs after infected.

• Parasites, Hosts and Diseases
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• 제51권2호
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• pp.213-218
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• 2013

Malaria is a parasitic infection caused by Plasmodium species. Most of the imported malaria in Korea are due to Plasmodium vivax and Plasmodium falciparum, and Plasmodium ovale infections are very rare. Here, we report a case of a 24-year-old American woman who acquired P. ovale while staying in Ghana, West Africa for 5 months in 2010. The patient was diagnosed with P. ovale malaria based on a Wright-Giemsa stained peripheral blood smear, Plasmodium genus-specific real-time PCR, Plasmodium species-specific nested PCR, and sequencing targeting 18S rRNA gene. The strain identified had a very long incubation period of 19-24 months. Blood donors who have malaria with a very long incubation period could be a potential danger for propagating malaria. Therefore, we should identify imported P. ovale infections not only by morphological findings but also by molecular methods for preventing propagation and appropriate treatment.