Rapid diagnosis of scrub typhus by a passive hemagglutination assay using recombinant 56-kilodalton polypeptides

Kim, Ik-Sang; Seong, Seung Yong; Woo, Sang-Gyu; Choi, Man-Sik; Kang, Jae-Seung; Chang, William

doi:10.1128/jcm.31.8.2057-2060.1993

Cited by 44 publications

(50 citation statements)

References 24 publications

(21 reference statements)

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“…PHA was performed using a Genedia TsuTsu PHA II test kit (GreenCross SangA, Yongin city, Kyunggi-do, Korea). This kit is currently used to quantitatively and qualitatively analyse antibodies against O. tsutsugamushi in human sera; it employs sheep red blood cells (RBCs) sensitized to recombinant 56-KDa protein from a Boryoung serovariant of O. tsutsugamushi specific for Korea, as well as the Karp and Gilliam serovariants of O. tsutsugamushi [9].…”

Section: Methodsmentioning

confidence: 99%

A serosurvey of Orientia tsutsugamushi from patients with scrub typhus

Kim¹,

Lee²,

Bäck³

et al. 2010

Clinical Microbiology and Infection

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Many countries where scrub typhus is endemic use their own cutoff values for antibody titres to differentiate between cured cases and current infections. To establish an antibody titre cutoff value, one needs to investigate the seroprevalence in endemic areas, and the duration of the increase in titre after complete cure. We conducted a follow-up study of anti-Orientia tsutsugamushi antibody titres using indirect immunofluorescence assays (IFA) and passive haemagglutination assays (PHA) in patients with scrub typhus. After the onset of symptoms, IgM antibody titres increased gradually over 2-3 weeks, peaked at about 4 weeks, and started to decrease rapidly between 4 and 5 weeks. At 1-year follow-up, the median IgM value was 1:10. Out of 77 patients who were tested at that time, 36 (47%) had IgM titres > or =1:20, and none had titres exceeding 1:80. Over the first 2 weeks, IgG antibody titres increased sharply, peaked at about 4 weeks and decreased rather gradually thereafter, with a median titre of 1:128 maintained up to the 18th month. At 1-year follow-up, five out of 77 patients (6.5%) had titres > or =1:1,024 and 57% had titres > or =1:128. Based on these results, a cutoff value of > or =1:160 for IgM antibody should differentiate between previous and current infections in endemic areas such as Korea and Japan, where scrub typhus occurs mainly in the autumn.

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Section: Methodsmentioning

confidence: 99%

A serosurvey of Orientia tsutsugamushi from patients with scrub typhus

Kim¹,

Lee²,

Bäck³

et al. 2010

Clinical Microbiology and Infection

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“…56-kDa protein, which was predicted to be hydrophobic and embedded in the membrane and showed no reactivity with human IgG and IgM antibodies (23). Previous reports have shown that 56-kDa protein was always produced in the form of inclusion body in E. coli (15)(16)(17)(18)(19)(20). However, the recombinant protein was highly soluble in our study.…”

Section: Discussionmentioning

confidence: 52%

“…Recent investigations suggested that the major outer membrane 56-kDa protein is a protective antigen that can be produced as a suitable recombinant protein for a diagnostic reagent purpose (15). Kim et al used a recombinant 56-kDa protein from the Boryong strain fused with maltose binding protein as the antigen in ELISA and a passive hemagglutination test for scrub typhus (17,18). Ching and colleagues have developed a rapid immunochromatographic flow test to detect the anti-O.…”

mentioning

confidence: 99%

Prokaryotic expression and immunogenicity of 56‐kDa protein of Orientia tsutsugamushi strain Karp

Wang

Chen

Qiang

et al. 2012

Microbiology and Immunology

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To express the 56‐kDa protein of O. tsutsugamushi strain Karp, this protein gene was cloned into pET30a(+) before transforming into host bacteria, E. coli Rossetta. Specificity of the recombinant protein was assessed by ELISA using rabbit sera against common members of the order Rickettsiae and 10 other pathogenic bacteria. After IPTG induction, SDS‐PAGE analysis of isolated protein demonstrated a band at approximately 46‐kDa. Western blot and mass spectrometry analysis proved that the recombinant protein was expressed successfully. Specificity analysis demonstrated that all sera were negative, except sera against O. tsutsugamushi strains TA763, TH1817 and Kato, B. quintana, A. phagocytophilum, E. chaffeensis and B. bacilliformis. The purified protein was used to immunize BALB/c mice and polyclonal antisera were harvested. By examination of IFA and ELISA, the highest titer of the polyclonal antibodies reaches 1:1600. The recombinant 56‐kDa protein in the study is valuable for developing a simple and rapid diagnostic test and vaccine for O. tsutsugamushi.

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“…When the mice became sick, their spleens were aseptically removed, minced and inoculated onto a mouse L cell culture. When about 80% of the cells were infected as confirmed by the IFA test, the cells were harvested and the organisms purified by Percoll density-gradient centrifugation as previously described (7,8,12 typhus including acute fever and skin rashes. A PHA test using rickettsial 56 kDa protein antigen was performed as previously described (8) and the patient's antibody titer was 1:640.…”

Section: Methodsmentioning

confidence: 99%

“…In Korea, scrub typhus is one of the main causes of acute febrile illness, especially in rural areas. The disease has usually been diagnosed serologically by IFA or passive hemagglutination assay (PHA) (1,8), and treatment has been by antibiotics. Because high incidence rate of scrub typhus in the fall harvesting season results in a significant loss of the labor force in rural areas, proper prevention of the disease is desirable.…”

mentioning

confidence: 99%

Isolation of a New Orientia tsutsugamushi Serotype

Seong

Park

Kim

et al. 1997

Microbiology and Immunology

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Orientia tsutsugamushi, the etiological agent of scrub typhus, is an antigenically diverse organism and many serologically distinct strains have been identified. The 56 kDa protein of O. tsutsugamushi, a major protein in the outer membrane, has been thought to be responsible for this antigenic variability. A strain of O. tsutsugamushi isolated in Korea cross-reacted with both Gilliam strain-specific and Karp strain-specific monoclonal antibodies. When its 56 kDa protein gene was cloned and analyzed, its sequence showed variation especially between 1,200 and 1,250 bp, showing that this isolate is a new O. tsutsugamushi strain.

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Rapid diagnosis of scrub typhus by a passive hemagglutination assay using recombinant 56-kilodalton polypeptides

Cited by 44 publications

References 24 publications

A serosurvey of Orientia tsutsugamushi from patients with scrub typhus

A serosurvey of Orientia tsutsugamushi from patients with scrub typhus

Prokaryotic expression and immunogenicity of 56‐kDa protein of Orientia tsutsugamushi strain Karp

Isolation of a New Orientia tsutsugamushi Serotype

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