• Title/Summary/Keyword: Indirect immunofluorescent assay(IFA)

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Survey of antibody on Orientia Tsutsugamushi among wild rodents in Gyeongnam area and detection by nested polymerase chain reaction (경남지역 야생들쥐에서 Orientia tsutsugamushi에 대한 항체 조사 및 PCR에 의한 검색)

  • Hah, Dae-Sik;Kim, Young-Hoon;Park, Jung-Ung;Park, Jae-Kap;Kim, Chung-Hui;Ryu, Jae-Doo;Jong, Myung-Ho;Heo, Jung-Ho;Shu, Jong-Lip;Cho, Myung-Heui;Lee, Kuk-Cheon;Kim, Gon-Sup;Kim, Eui-Kyung;Kim, Jong-Shu
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.241-249
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    • 2004
  • As a part of epidemiologic investigation of tsutsugamushi disease, the wild rodents which were captured in Gyeongnam area were diagnosed with indirect immunofluorescent antibody assay (IFA) and Polymerase Chain Reaction (PCR) to find if they have an antibody against Orientia tsutsugamushi. The conclusion was drawn as followings. (1) The captured 58 wild rodents showed that the subspecies distribution of Apodemus agrarius was 86.2%, Microtus fortis was 8.6% and Crocidura lasiura was 5.2%. (2) The antibody positive rate of O. tsutsugamushi Gilliam, Karp, Karto and Boryong by IFA method was 32.0% in Apodemus agrarius among 50 wild rodents and 40.0% in Microtus fortis among 5 wild rodents, respectively. It was negative in the case of all the 3 Crocidura lasiura. (3) The antibody titers on Apodemus agrarius, Microtus fortis and Crocidura lasiura against Gilliam, Karp, Karto and Boryong were measured between 1:20 and 1:640. The antibody titer against each antigen was in the order Boryong>Gilliam>Karp. (4) O. tsutsugamushi was detected from the blood, spleen and kidney from the artificially infected mice by IFA and PCR. IFA showed the positive response between 3 and 18 days after inoculation. On the other hand, positive response was found from all the samples by PCR. (5) From PCR of the genomic DNA extracted from the blood, spleen and kidney samples of the captured wild rodents, Boryong-specific amplification product with size of 210 bp, which is particular in Boryong, was detected from spleen and kidney samples, but not detected in the blood. (6) Boryong-specific amplification product was detected from spleen and kidney samples which were obtained at 3, 6, 12, 18 and 24 days after the infection with Boryong. But, it wasn't detected from the uninfected samples. (7) From PCR of spleen and kidney samples of the captured wild rodents, it was found that positive rate of O. tsutsugamushi in Apodemus agrarius and Microtus fortis were 25.0% (4/16) and 20.0% (1/5), respectively. From the above results, it can be concluded that Apodemus agrarius resided in Gyeongnam area carried O. tsutsugamushi and PCR method might be a simple, precise, rapid and useful diagnostic tool than IFA for the diagnosis of O. tsutsugamushi.

Prevalence of Antibodies to Human Herpesvirus 8 in Children (소아의 항 Human Herpesvirus 8 항체 양성률)

  • Han, Tae Hee;Chung, Ju Young;Kim, Sang Woo
    • Pediatric Infection and Vaccine
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    • v.12 no.2
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    • pp.108-113
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    • 2005
  • Purpose : Human herpsevirus 8(HHV-8), a gamma herpsevirus, was initially identified from Kaposi sarcoma(KS) lesions and has been known to be associated with several malignancies including Kaposi sarcoma. HHV-8 seroprevalence is variable by different geographic areas and populations. The prevalence of HHV 8 infection in Korean children is unclear. So, we investigated the prevalence of HHV-8 specific antibodies in healthy children in Seoul, Korea. Methods : Sera were obtained from 112 children(age 1~15 years, 64 males and 48 females) who visited our hospital for routine health checkup and used for investigating sero-prevalence of anti-HHV-8 antibodies. An indirect immunofluorescent assay was used to detect the IgG antibodies to the lytic viral antigen(Biotrin, Dublin, Ireland). A peptide mix ELISA kit was used to detect the IgG antibodies to peptides specific for HHV-8 open reading frame (ORF)(Biotrin, Dublin, Ireland). Results : Of 112 children, 4 children younger than 6 years of age were seropositive to HHV-8[all 4(3.5%) were positive by IFA and 2(1.8%) were positive by ELISA]. Conclusion : These results suggest that the prevalence of antibody to HHV 8 in children in Korea is very low.

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Immunohistoehemical Observation on the Antigens Inducing IgG and IgM Antibodies against Sparganum (IgG와 IgM 항체를 유도하는 sparganum의 항원에 관한 면역조직화학적 및 전기영동에 의한 연구)

  • 김창환;최완성
    • Parasites, Hosts and Diseases
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    • v.29 no.4
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    • pp.339-354
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    • 1991
  • Localization and characterization of the antigenic components of sparganum which induced IgG and IsM antibodies in the host were studied by immunohistochemical techniques and SDS-PAGT and Western blotting. The antigen recognized by IgG antibody of rats or mice which were immunised by infection or injection of crude extracts of metacestodes of Spirometra erinacei, was located in the parenchyme of sparganum, especially at the cortex and around the calcareous corpuscles. The immunoreaction was demonstrated not only in the encysted fibrous wall of host but around the arterioles or venules in the connective tissue of host. The antigen recognized by IgM antibody of rats or mice was also observed in the parenchyme of sparganum and in the connective tissue of host. By 5∼20% gradient SDS-PAGE and EIBT, we detected antigenic components by IgG and 1gG antibodies of the rat or mouse immunized by infection or injection of crude extract of spargana. Twenty-three antigenic bands from crude extracts of spargana were recognized by IgG antibody and 15 components by IgM antibody of immunized rats. Out of the bands recognized by IgG and IgM antibodies, 15 were cross-reacted each other. Twenty components of eBlcretory-secretory proteins from spargana were recognized by IgG, and 5 components by IgM antibody of immunized rats. By IgG and IgM antibodies of immunized mice, 16 components of crude extracts were recognized by IgG antibody and 9 components by IgM antibody. Twenty components of excretory-secretory preparation were recognized by IgG antibody and 5 components by IgM antibody. Thirteen components of crude extracts were cross-reacted by IgG antibody of rats and mice.

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