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Rickettsial infections and fever, Vientiane, Laos.

Rickettsial diseases have not been described previously from Laos, but in a prospective study, acute rickettsial infection was identified as the cause of fever in 115 (27%) of 427 adults with negative blood cultures admitted to Mahosot Hospital in Vientiane, Laos. The organisms identified by serologic analysis were Orientia tsutsugamushi (14.8%), Rickettsia typhi (9.6%), and spotted fever group rickettsia (2.6% [8 R. helvetica, 1 R. felis, 1 R. conorii subsp. indica, and 1 Rickettsia "AT1"]). Patients with murine typhus had a lower frequency of peripheral lymphadenopathy than those with scrub typhus (3% vs. 46%, p<0.001). Rickettsioses are an underrecognized cause of undifferentiated febrile illnesses among adults in Laos. This finding has implications for the local empiric treatment of fever.

Adolescent↗

Rapid and simple identification of Orientia tsutsugamushi from other group rickettsiae by duplex PCR assay using groEL gene.

In this study, two new duplex PCR methods based on the groEL gene were developed and investigated for the diagnosis of rickettsiae. The first duplex PCR assay amplified the 229-bp and the 366-bp DNAs of 6 strains including typhus group (TG) and spotted fever group (SFG) rickettsiae, and 5 scrub typhus group (STG) rickettsiae, respectively. The second duplex PCR assay amplified the 397-bp and the 213-bp DNAs of 6 Rickettsia strains and 5 STG strains. These duplex PCR methods could simultaneously perform the rapid identification of rickettsiae and the differential diagnosis of STG and other group rickettsiae in a single reaction.

Bacterial Typing Techniques↗

Short report: prospective evaluation of a multi-test strip for the diagnoses of scrub and murine typhus, leptospirosis, dengue Fever, and Salmonella typhi infection.

A multi-test strip dotblot immunoassay for the diagnosis of typhoid fever, scrub typhus, murine typhus, dengue virus infection and leptospirosis was evaluated in Thai adults presenting to hospital with acute, undifferentiated fever. The kit gave multiple positive test results in 33 of 36 patients with defined infections and was therefore not a useful admission diagnostic tool.

Adolescent↗

MITES (FAMILY TROMBICULIDAE) PARASITIZING BIRDS MIGRATING FROM AFRICA TO EUROPE.

The mechanisms of dissemination of arthropod-borne human and animal pathogens are of considerable interest to the epidemiologist, veterinarian and biologist. Birds which are hosts to such pathogens and their arthropod vectors could transport them over long distances during their spring and autumn migratory flights.In April 1961, birds migrating from Africa to Europe were collected in south-western Spain and examined for ectoparasites and antibodies to arboviruses. Fully engorged larvae of two species of trombiculid mites unknown in Europe (genera Neoschoengastia and Blankaartia) but found in Africa were collected from two of the migrating birds (redstart and little bittern), suggesting that the birds were carrying the mites from Africa to Europe.Trombiculid mites are the proven vectors of scrub typhus; they have also been implicated in the transmission of human haemorrhagic nephroso-nephritis. The finding of the mite larvae on migrating birds is therefore of some epidemiological interest and underlines the importance of obtaining more data on the dispersal of trombiculids by migrating birds.

Africa↗

Rickettsioses in Australia.

Australia, an island continent in the southern hemisphere, has a range of rickettsial diseases that include typhus group rickettsiae (Rickettsia typhi), spotted fever group rickettsiae (R. australis, R. honei), scrub typhus group rickettsiae (R. tsutsugamushi), and Q fever (C. burnetii). Our knowledge of Australian rickettsiae is expanding with the recognition of an expanded range of R. honei (Flinders Island spotted fever) to Tasmania and southeastern mainland Australia (not just on Flinders Island), and the detection of a new SFG species (or subspecies), tentatively named "R. marmionii" in the eastern half of Australia. This rickettsia causes both acute disease (7 cases, recognized so far) and is also associated (as a "R. marmionii" bacteriaemia) with patients having a chronic illness. The significance of the latter is under investigation. It may be a marker of autoimmune disease or chronic fatigue in some patients.

Animals↗

Differentiation of rickettsiae by groEL gene analysis.

The nucleotide sequences (534 to 546 bp) of the groEL gene, which encodes the 60-kDa heat shock protein GroEL, from 15 rickettsial strains were determined and compared. In the phylogenetic tree created by the unweighted pair group method with arithmetic averages and the neighbor-joining method, rickettsial strains could be distinguished from Ehrlichia strains. Five spotted fever group strains, four typhus group strains, and six scrub typhus group (STG) strains were differentiated as distinct entities. Unlike gltA and ompA gene analyses, differentiation between members of the genus Rickettsia and the STG rickettsiae by groEL gene analysis was possible. In comparison with 16S rRNA gene analysis, the groEL gene has a higher degree of divergence among the rickettsiae. We therefore successfully developed rapid differentiation methods, PCR-restriction fragment length polymorphism analysis and a species-specific PCR, based on the groEL gene sequences. Four Korean isolates were identified by these methods and groEL gene analysis. The results suggest that the groEL gene is useful for the identification and characterization of rickettsiae.

Bacterial Typing Techniques↗

Preliminary evaluation of the INDX DIP-S-TICKS with positive rickettsial samples in Malaysia.

Forty-four serum samples of various reactivities to rickettsial antigens demonstrated by the indirect immunoperoxidase technique were tested with INDX Dip-S-Ticks (INDX Integrated Diagnostics Inc., USA) Kit for the detection of tick borne diseases. The kit utilised Rickettsia rickettsii the causative agent of Rocky Mountain spotted fever (RMSF) as antigens. The samples positive for endemic typhus were also tested against R. typhi, the agent for endemic typhus by the same method. The aim of this study was to determine the extent of cross-reactivity of R. rickettsii with rickettsial infections in Malaysia. Nine out of 12 tick typhus, 4 out of 10 scrub typhus and 4 out of 12 endemic typhus samples cross reacted with R. rickettsii. Ten out of 12 endemic samples were positive with R. typhi by the same method. From the study, we concluded that the INDX Dip-S-Ticks Kit can be used as a rapid screening test to detect endemic and tick-borne rickettsial infections in Malaysia but a second serological test is strongly recommended on all weakly reactive cases.

Animals↗

A survey of scrub and murine typhus in the Ancol section of Jakarta, Indonesia.

Rickettsia tsutsugamushi was isolated from L. (L.) arenicola chiggers and three species of rats in an area of scrub and sedge along the Bay of Jakarta. This is the only finding in Indonesia of a cycle of the agent of scrub typhus associated with L. (L.) arenicola. A serologic survey of nearly 300 persons living in two kampungs near the site at which rickettsiae were recovered revealed one individual with antibodies to R. tsutsugamushi. Murine typhus, with a seropositivity rate of 6.5%, may be endemic at low levels.

Animals↗

Rickettsial meningitis and encephalitis.

Nine of 72 patients with scrub typhus and three of 137 with murine typhus presented with meningitis and/or encephalitis syndromes. Focal neurologic signs were rare, and cerebrospinal fluid profiles were similar to those of leptospirosis and viral and tuberculous meningitis. One patient had papilledema, and another had cerebellitis. Other major organ involvement (renal, liver, or lungs) occurred in five patients. One patient died and four spontaneously recovered, while the conditions of the rest responded well to either chloramphenicol or doxycycline. Scrub and murine typhus should be included in the differential diagnoses of aseptic meningitis and encephalitis in patients exposed to endemic areas, especially when accompanied by renal insufficiency and/or jaundice. They are treatable forms of virallike meningoencephalitis.

Adolescent↗

Scrub and murine typhus in children with obscure fever in the tropics.

Between October, 1985, and February, 1987, 28 (8.7%) cases of scrub and murine typhus were diagnosed among 320 children with greater than or equal to 1 week history of obscure fever. Scrub typhus is a rural disease and characterized by fever, tachypnea and hepatosplenomegaly. Skin rash was rare and eschar was absent. Four patients had pneumonia and two had meningitis. Murine typhus, more an urban disease, was milder and half the patients presented exclusively because of night fever. Slightly enlarged liver and skin rash were the only significant physical signs. Lacking the classical textbook presentations, both rickettsioses often were missed or diagnosed as enteric fever. Recognition is important because patients with either disease respond well to treatment with chloramphenicol or doxycycline.

Child↗

A scanning and transmission electron microscopic examination of Rickettsia tsutsugamushi-infected human endothelial, MRC-5, and L-929 cells.

Monolayers of primary human endothelial cells were infected with the Karp strain of Rickettsia tsutsugamushi and examined by scanning and transmission electron microscopy. The results were compared with those obtained with similarly infected L-929 and MRC-5 cells and with uninfected cells of all three types. The rickettsiae grew to slightly higher titers in the human endothelial cells. Transmission electron microscopy revealed significant changes in the host cell organelles; a reduction in ribosome-coated endoplasmic reticulum and in Golgi activity, swelling of mitochondria, and an increase in vacuolation within the cytoplasm. Since human endothelial cells are known to retain their in vivo structural and functional qualities when cultured in vitro, it is likely that these effects are similar to those which occur during the infectious process in human scrub typhus.

Animals↗

Psoralen photochemical inactivation of Orientia tsutsugamushi in platelet concentrates.

BACKGROUND: The risk of transfusion transmission of disease has been reduced by the combination of predonation questions and improved transfusion-transmitted disease assays, but the risk is still present. This study was conducted to determine if psoralen photochemistry could inactivate an obligate intracellular bacterium, with documented potential for transfusion, in PCs to further improve safety. STUDY DESIGN AND METHODS: PCs were inoculated with MNCs infected with Orientia tsutsugamushi. The concentrates were treated with amounts ranging from 0.86 to 138 micromol per L of 4'-(aminomethyl)-4,5',8-trimethylpsoralen hydrochloride (AMT) combined with a constant long-wave UVA light (320-400 nm) exposure of 5 J per cm(2). The effects of photochemical treatment were analyzed by using a mouse infectivity assay along with in vitro testing by PCR, indirect fluorescence antibody, direct fluorescence antibody, and Giemsa staining. RESULTS: AMT, at 0.86 micromol per L or more, combined with UVA light of 5 J per cm(2), inactivated O. tsutsugamushi that contaminated PCs. The PCs that did not receive the combined treatment caused infection. CONCLUSIONS: The psoralen AMT, in conjunction with UVA light exposure, effectively abolished the infectivity of PCs deliberately contaminated with the scrub typhus organism O. tsutsugamushi, as tested in a mouse infectivity assay.

Animals↗

Susceptibility of Rickettsia tsutsugamushi Gilliam to gamma interferon in cultured mouse cells.

Recombinant rodent gamma interferon (IFN-gamma) inhibited the infection of cultured BALB/3T3 mouse fibroblasts by Rickettsia tsutsugamushi Gilliam, apparently mainly by clearance of intracellular rickettsiae. No significant effect on rickettsial entry into the cells was noted; IFN-gamma was toxic to infected cells, as measured by the capacity of treated, infected cells to attach to the surfaces of culture vessels. In a small proportion of IFN-gamma-treated cells, rickettsial replication appeared to persist at normal levels. A fraction (28%) of rickettsiae clonally isolated from cultures treated with IFN-gamma was resistant to IFN-gamma-mediated inhibition, but four serial passages of these resistant clones in the absence of additional IFN-gamma resulted in the loss of resistance. In several respects, therefore, the IFN-gamma-mediated inhibition of scrub typhus rickettsiae in cultured fibroblasts was similar to that reported for Rickettsia prowazekii.

3T3 Cells↗

Efficiency of Leptotrombidium chiggers (Acari: Trombiculidae) at transmitting Orientia tsutsugamushi to laboratory mice.

Thirteen different laboratory colonies of Leptotrombidim chiggers [L. chiangraiensis Tanskul & Linthicum, L. deliense Walch and L. imphalum (Vercammen-Grandjean &Langston)] were evaluated for their ability to transmit Orientia tsutsugamushi (Hyashi) to mice. Of 4,372 transmission attempts using individual chiggers from all 13 colonies, 75% (n = 3,275) successfully infected mice. Transmission rates for the individual chigger colonies ranged from 7 to 80%. Increasing the number of chiggers that fed on a given mouse generally increased transmission rates. Transmission of O. tsutsugamushi to mice by different generations (F1-F11) of certain chigger colonies was stable; however, transmission rates varied greatly in other colonies. Transmission rates (both vertical and horizontal) of several L. changraiensis colonies and the L. deliense colony were the highest, suggesting that these colonies may be useful for the development of a chigger-challenge model that can be used to evaluate the efficacy of candidate scrub typhus vaccines or therapeutic agents in laboratory mice.

Animals↗

[Investigation on rodents' natural infection of Orientia tsutsugamushi in some areas of Inner Mongolia and Xinjiang, China].

OBJECTIVE: To investigate rodents' natural infection of Orientia tsutsugamushi (Ot) in some areas of Inner Mongolia and Xinjiang, China. METHODS: DNAs were extracted from spleens of the captured mice and nested-polymerase chain reaction (nPCR) technique was used to detect the Ot-Sta56 gene. Six positive samples were sequenced and analyzed by Clustal X (5.0) and DNA Club software. RESULTS: A total of 90 rodents were captured in Inner Mongolia, and the overall prevalence of Ot was 6.67%. There was no significant difference in infection rates among the positive rodents species. 20 rodents were captured in Xinjiang, and the prevalence of Ot was 5.00%. The geographical difference in infection rates was not statistically significant between Inner Mongolia and Xinjiang. 9 rodents were captured in farmlands of Inner Mongolia and Xinjiang but there was no positive samples found. 101 rodents were captured in grasslands, and the prevalence of Ot was 6.93%. The Sta56 gene nucleotide sequence homology to Karp strain of N59 (from Microtus maximowiczii), N69 (from Cricetulus barabensis) and X33(from Cricetus cricetus) was 99%. The sequence homology to Taitung-2 strain and TW461 strain of N65 (from C. barabensis) was 94%, and the sequence homology to Taitung-2 strain and TW461 strain of N88(from Apodemus agrarius) was also 94%. The sequence homology to Oishi strain of N90 (from A. agrarius) was 96.00%. CONCLUSION: Our findings indicated that infections of Ot did exist in rodents captured from Inner Mongolia and Xinjiang. The genotypes of Ot in Inner Mongolia and Xinjiang were quite complex, with some of them belonged to Karp type, and the others belonged to Taitung-2, TW461 and Oishi types which providing evidence for further investigation on the scrub typhus fuci in the two areas.

Animals↗

Detection of Rickettsia tsutsugamushi specific DNA from the lymphocyte of patients by polymerase chain reaction.

Polymerase chain reaction was used to detect Rickettsia tsutsugamushi specific DNA from the lymphocytes of patients in the acute phase of the disease. The primers used in the PCR were based on the nucleotide sequences of gene encoding of the 56 kDa antigen of the Gilliam strain. The PCR product is a 78 bp fragment which can be hybridized by the 78 bp DNA probe of the Gilliam strain. Comparison of the results obtained by polymerase chain reaction, immunofluorescent assay and cell culture suggests that the polymerase chain reaction method is the most sensitive one among the three in diagnosis of scrub typhus.

Acute-Phase Reaction↗

Invasion and intracellular growth of Rickettsia tsutsugamushi.

Intracellular multiplication of Rickettsia tsutsugamushi, a causative agent of scrub typhus, was examined by electron microscopy of specimens prepared at various time intervals after infection of in vitro cultured cells. The sequential morphological growth cycle of the microorganism is presented diagrammatically.

Animals↗