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PubMed2002

Expression of recombinant proteins of Orientia tsutsugamushi and their applications in the serodiagnosis of scrub typhus.

In this study, recombinant proteins that encompassed the AD I-AD III regions of 56 kDa immunodominant gene of 2 Orientia tsutsugamushi (OT) serotypes; Gilliam and TA763 were expressed in Escherichia coli. Both recombinant proteins exhibited serologic cross-reactivity with the rabbit antisera against various OT serotypes, as evaluated by enzyme-linked immunosorbent assay (ELISA), but not against other rickettsial species, including Rickettsia typhi, R. prowazekii and TT118 SFG rickettsiae. The feasibility of using the recombinant proteins as a diagnostic reagent was further evaluated by ELISA using sera from blood donors and scrub typhus patients. The results suggested a higher affinity of the antihuman IgM than IgG with both recombinant proteins. The IgM ELISA findings were agreeable with the results of indirect immunoperoxidase (IIP) assay especially with sera of high antibody (1:1600). However, more than one antigen are probably needed for development of an effective assay for serodiagnosis of scrub typhus in endemic areas.

Animals↗
PubMed1997

Rudolf Virchow, not just a pathologist: a re-examination of the report on the typhus epidemic in Upper Silesia.

Rudolf Virchow (1821-1902) is mainly remembered as the "father" of cellular pathology; however, he was not just a pathologist. His contributions to anthropology, archeology, ethnography, and history, as well as his involvement in epidemiology, public health, and politics, portray a man with multiple interests, deeply engaged in the controversies of his time. In his Report on the Typhus Epidemics of Upper Silesia of 1848, the young Virchow reveals himself to be a self-assured pathologist, although his postmortem examinations failed to shed much light on typhus. Despite of his shortcomings and biases, Virchow's genius is revealed in his deep appreciation of the importance of the total physical, socio-cultural, economic, and political background of epidemic diseases. One can discern in the Report the making of Virchow, the politician and statesman who will contribute to the modernization of Germany's public health, and of the physician-scholar and physician-citizen who, despite of his shortcomings and militancy, continues to inspire and challenge us today.

Disease Outbreaks↗
PubMed2001

Identification of the target cells of Orientia tsutsugamushi in human cases of scrub typhus.

Orientia tsutsugamushi is the etiologic agent of scrub typhus, a chigger-borne zoonosis that is a highly prevalent, life-threatening illness of greatest public health importance in tropical Asia and the islands of the western Pacific Ocean. The target cell of this bacterium is poorly defined in humans. In this study, O. tsutsugamushi were identified by immunohistochemistry using a rabbit polyclonal antibody raised against O. tsutsugamushi Karp strain in paraffin-embedded archived autopsy tissues of three patients with clinical suspicion of scrub typhus who died during World War II and the Vietnam War. Rickettsiae were located in endothelial cells in all of the organs evaluated, namely heart, lung, brain, kidney, pancreas, and skin, and within cardiac muscle cells and in macrophages located in liver and spleen. Electron microscopy confirmed the location of rickettsiae in endothelium and cardiac myocytes.

Adult↗
PubMed1998

Recovery and viability of Orientia tsutsugamushi from packed red cells and the danger of acquiring scrub typhus from blood transfusion.

BACKGROUND: The purpose of this study was to determine whether infective Orientia tsutsugamushi, the etiologic agent of scrub typhus, could survive normal blood banking processing and storage procedures. STUDY DESIGN AND METHODS: Mononuclear cells isolated from whole blood by density gradient centrifugation were inoculated with O. tsutsugamushi, Karp strain. Infection of the mononuclear cells was confirmed by Giemsa stain, direct fluorescent antibody assay, and polymerase chain reaction using primers specific for the groESL operon of O. tsutsugamushi. The quantity of rickettsial particles in each preparation was determined by direct counts from the Giemsa-stained preparations. Infected mononuclear cells were returned to their respective aliquots of packed red blood cells, which were then either stored at 4 degrees C or glycerolized and frozen at -70 degrees C. RESULTS: Rickettsiae survived up to 10 days (but not 30 days) of refrigerated storage and 45 days of frozen storage, as determined by inoculation of mice with 0.5-mL aliquots of the blood components. Infection of the mice was determined by illness, death, direct fluorescent antibody assay of peritoneal smears, polymerase chain reaction of blood, and enzyme-linked immunosorbent assay detection of antibodies in plasma. CONCLUSION: Because the quantity of rickettsiae injected into the mice was comparable to the quantity reported in the literature for human blood during natural infections, scrub typhus could present a risk in blood collected from donors in endemic areas. This may especially be true, because people can be rickettsemic before illness, after successful antibiotic treatment, and chronically after resolution of disease.

Animals↗
PubMed2001

[Fever and calf tenderness after travel in Thailand: murine typhus and deep vein thrombosis].

HISTORY AND CLINICAL FINDINGS: A 37-year-old German fell ill with fever and diffuse headaches 3 weeks after flying to Thailand for a holiday. Because of increasing sickness he began his return (13 h by train then 11 h by plane). Besides a temperature of 39.3 degrees C he presented with a unproductive cough and pain in his left calf. On examination there were no other pathological findings than a borderline tachycardia, mild hypotension and signs of dehydration. INVESTIGATIONS: Laboratory tests showed elevated inflammatory parameters (CRP 77.8 mg/l, ESR 36 mm), normal range of leucocytes with obvious shift to the left, mild thrombocytopenia and elevated liver enzymes. The Weil-Felix-reaction revealed a high antibody titre against OX 19, the IFT for Rickettsia typhi a significant increase of IgM with seroconversion for IgG. Duplex ultrasonography and phlebography confirmed a phlebothrombosis of the left upper thigh. DIAGNOSIS: Murine typhus and deep vein thrombosis of the left upper leg. TREATMENT AND COURSE: Suspecting typhoid fever with ciprofloxacin was started. In less then 2 days the patient became afebrile and recovered. The deep vein thrombosis was treated with low-molecular-weight heparin (Tinzaparin) overlapping with phenprocoumon. CONCLUSION: In patients with fever after travel to tropical or subtropical areas (including southern Europe) the diagnosis of murine typhus should be considered, especially if concomitant with thromboembolic events. The Weil-Felix-reaction is a helpful screening test supplemented by Rickettsia specific serologic tests. Overall a higher risk of thrombosis should be taken into account after long sedentary travel.

Adult↗
PubMed1994

[Rickettsiosis of the typhus fever type. Its joint appearance with a left-ventricular thrombus and single-vessel coronary disease].

A 44-year-old German fell ill in Libya, where he had been living for 10 years, with high fever, rigor and a nonitching centrifugally spreading macular rash, which had spared the head, hands and soles. In addition, a systolic cardiac murmur was heard. The Weil-Felix reaction had a titre rising within 3 days from 1:160 to 1:640, confirming the diagnosis of rickettsial disease, the total clinical picture indicating typhus. On treatment with chloramphenicol (1 g three times daily i.v.) the fever subsided within 5 days. On the ninth day treatment was changed to oral doxycyclin, 200 mg daily for 3 weeks. Echocardiography surprisingly revealed a floating thrombus, about 4 x 8 cm, attached to the hypo- and even akinetic apex of the left ventricle. In addition there was single-vessel coronary disease. Since the segmental contraction abnormality persisted after the typhus had been cured, a causal connection with the rickettsial disease is unlikely. The thrombus was removed at the time of a aortocoronary bypass operation: his course has been unremarkable since then.

Adult↗
PubMed1993

[Interstitial nephritis in typhus].

A 58-year-old man developed continuous fever of around 40 degrees C and a macular rash on the trunk and limbs one week after an eight-day visit to the island of Rhodos. Typhus was diagnosed on the basis of the clinical findings, the characteristic rash and a rise in antibody titre against Rickettsia prowazekii and mooseri (1:1280). The fever subsided and the patient's general state markedly improved within three days of starting antibiotic treatment with doxycycline. Five days after hospital admission renal function deteriorated, serum creatinine rising from 1.7 to maximally 8.7 mg/dl. Renal biopsy on the eighth day of illness revealed, in concordance with the diagnosis of a rickettsial infection, acute granulomatous interstitial nephritis. Doxycycline administration was continued for a total of 18 days and renal function improved. Two weeks after discharge serum creatinine had fallen to 1.8 mg/dl and to 1.2 mg/dl after 6 months. The antibody titre against Rickettsia prowazekii had fallen to 1:20 about 12 weeks after discharge. Early antibiotic treatment of typhus is of importance to avoid irreversible renal damage.

Antibodies, Bacterial↗
PubMed1994

Transmission cycle of murine typhus in Greece.

A field study, based on a follow up of the 49 human cases of the disease that occurred on the island of Evia in 1985, has established the transmission cycle of murine typhus in Greece for the first time. In 1993, two types of traps were used to catch 53 rats, all of them Rattus norvegicus, in the localities where the cases had occurred. Some 300 fleas, all of them probably Xenopsylla cheopis, and a few ticks and mites were collected from the rats. Eight of the fleas were found positive for Rickettsia typhi by PCR and anti-R. typhi antibodies were found in the sera of 48 (91%) of the rats at titres between 1:8 and 1:524,288 and in 22 (42%) at titres > 1:256. The commensal rat population in the general area of study was both ubiquitous and abundant. Murine typhus is obviously endemic in the area and probably of much greater public health significance than the number of reported cases indicates.

Animals↗
PubMed2006

Scrub typhus imported to Scandinavia.

We present the first 3 cases of confirmed scrub typhus imported to Scandinavia. The patients were infected in Thailand, Laos and Sri Lanka, respectively. Treatment with ciprofloxacin and doxycycline lead to rapid cure in 2 of the patients, while the third patient, who was not treated, had a prolonged convalescence. Specific antibodies against Orientia tsutsugamushi, the causative agent of scrub typhus, were detected in all 3 cases.

Adult↗
PubMed2000

Multiple organ failure complicating probable scrub typhus.

This report describes a case of life-threatening acute respiratory distress syndrome with multiple organ failure complicating probable scrub typhus. Favorable outcome was associated with fluoroquinolone therapy. Scrub typhus should be suspected in travelers returning from Southeast Asia presenting with unexplained respiratory manifestations.

Adult↗
PubMed1998

Performance of a dot blot immunoassay for the rapid diagnosis of scrub typhus in a longitudinal case series.

A rapid dipstick test for scrub typhus was prospectively evaluated in Chiangrai, northern Thailand. Sera from 162 patients with fever of unclear etiology were tested by a dot blot immunoassay using two different antigen concentrations. Dipsticks coated with lower concentration of antigen lacked sensitivity compared with the indirect immunoperoxidase test. Dipsticks with higher antigen concentration had increased sensitivity that was equivalent to that of the immunoperoxidase test. By increasing the antigen concentration on the dipstick, sensitivity increased from 67% to 100%, positive predictive value increased from 90% to 93%, and negative predictive value rose from 92% to 100%. The specificity of both antigen concentrations was 98%. This study establishes that scrub typhus can be confirmed serologically by use of a dipstick assay and that serodiagnosis can be effectively tailored to a target population.

Adult↗
PubMed1995

Short-course doxycycline treatment versus conventional tetracycline therapy for scrub typhus: a multicenter randomized trial.

To assess the clinical efficacy of short-course doxycycline in the treatment of scrub typhus, we compared conventional 7-day tetracycline therapy with 3-day doxycycline therapy in 116 patients. Patients were randomized to receive either tetracycline (500 mg four times daily; n = 50) or doxycycline (100 mg twice daily; n = 66) and were followed for 4 weeks after the completion of treatment. The cure rate was 100% in the tetracycline group and 93.9% in the doxycycline group (P > .05). The two groups did not differ significantly in terms of the interval required for defervescence or for the alleviation of symptoms. There were no relapses in either group. These data suggest that 3-day doxycycline therapy is as effective as conventional 7-day tetracycline therapy for the cure of scrub typhus and the prevention of relapses.

Adult↗
PubMed1996

Liver involvement in murine typhus.

Liver involvement was retrospectively evaluated in 137 patients with murine typhus. Fifteen (10.9%) were jaundiced. One patient had been subjected to cholecystectomy after misdiagnosis of acute cholecystitis. Serum aminotransferase levels were abnormal in 48/52 measurements, and there were elevations of > 5-fold in 14 patients. Liver biopsies and/or necropsies from four jaundiced patients showed portal tract and sinusoidal infiltrates, cloudy swelling/and necrosis of the hepatocytes and occasional pseudogranuloma formation. There were striking mitoses even in the early stage, suggesting rapid hepatocellular regeneration. Haemolytic diseases (G6PD deficiency or haemoglobinopathies), alcoholism, and a second infection probably also contributed to the pathogenesis of jaundice in murine typhus. This rickettsiosis should be included among differential diagnoses of acute hepatitis in patients exposed to areas endemic for Rickettsia typhi.

Acute Disease↗
PubMed1987

Serologic evidence of acute murine typhus infection in a patient with culture-negative endocarditis.

A patient with culture-negative infective endocarditis is presented in whom detailed serological studies were indicative of acute infection with murine typhus. The patient had aortic and mitral regurgitation with congestive heart failure and typical peripheral manifestations of subacute endocarditis, but no documented fever. Aortic and mitral valve replacement surgery and a 6-week course of doxycycline therapy produced a clinical cure in this patient, as well as a diagnostic fall in markedly elevated preoperative typhus indirect fluorescent antibody (IFA) and complement fixation (CF) titers. Serological studies were consistently negative for Q fever.

Acute Disease↗
PubMed2003

Scrub typhus and tropical rickettsioses.

PURPOSE OF REVIEW: Recent developments in molecular taxonomic methods have led to a reclassification of rickettsial diseases. The agent responsible for scrub typhus (Orientia tsutsugamushi ) has been removed from the genus Rickettsia and a bewildering array of new rickettsial pathogens have been described. An update of recent research findings is therefore particularly timely for the nonspecialist physician. RECENT FINDINGS: An estimated one billion people are at risk for scrub typhus and an estimated one million cases occur annually. The disease appears to be re-emerging in Japan, with seasonal transmission. O. tsutsugamushi has evolved a variety of mechanisms to remain viable in its intracellular habitat. Slowing the release of intracellular calcium inhibits apoptosis of macrophages. Subsets of chemokine genes are induced in infected cells, some in response to transcription factor activator protein 1. Cardiac involvement is uncommon and clinical complications are predominantly pulmonary. Serious pneumonitis occurred in 22% of Chinese patients. Dual infections with leptospirosis have been reported. Standardized diagnostic tests are being developed and attempts to improve treatment of women and children are being made. Of the numerous tick-borne rickettsioses identified in recent years, African tick-bite fever appears to be of particular importance to travellers. The newly described flea-borne spotted fever caused by Rickettsia felis may be global in distribution. SUMMARY: Rash and fever in a returning traveler could be rickettsial and presumptive doxycycline treatment can be curative. Recent research findings raise more questions than answers and should stimulate much needed research.

Animals↗
PubMed1995

Serological studies of the antigenic similarity between typhus group rickettsiae and Weil-Felix test antigens.

The sera from two patients with murine typhus reacted with whole cells of Rickettsia prowazekii, R. typhi, and Proteus vulgaris OX19, and with lipopolysaccharides (LPS) from the spotted fever group rickettsia strain TT-118 and P. vulgaris OX19 in the enzyme-linked immunosorbent assay. Sera from these patients reacted with ladder-like bands of LPS from R. prowazekii and R. typhi in the immunoblot, whereas the reactivity of these sera with LPS from P. vulgaris OX19 differed from each other. These results indicate that LPS from the typhus group rickettsiae and P. vulgaris OX19 contain similar epitopes.

Antibodies, Bacterial↗
PubMed1995

In vitro effectiveness of azithromycin against doxycycline-resistant and -susceptible strains of Rickettsia tsutsugamushi, etiologic agent of scrub typhus.

In an effort to find a potential alternative treatment for scrub typhus, we evaluated the effectiveness of the standard drug doxycycline and the new macrolide azithromycin against a doxycycline-susceptible strain (Karp) and a doxycycline-resistant strain (AFSC-4) of Rickettsia tsutsugamushi. The antibiotics were tested in an in vitro assay system in which infected mouse fibroblast cells (L929) were incubated for 3 days in various concentrations of the drugs. Rickettsial growth was evaluated by direct visual counts of rickettsiae in Giemsastained cells or by flow cytometry. Initial tests were conducted at the concentration of each antibiotic considered to be the upper breakpoint for susceptibility (16 micrograms/ml for doxycycline and 8 micrograms/ml for azithromycin). Growth of both Karp and AFSC-4 was strongly inhibited with both antibiotics, as measured by visual counts, although the percentage of cells infected with AFSC-4 in the presence of doxycycline was three times greater than the percentage of cells infected with Karp but was only 60% as great as the percentage of cells infected with Karp in the presence of azithromycin. Flow cytometry confirmed that rickettsial growth occurred in the absence of antibiotics, but it failed to detect it in the presence of high concentrations of either drug. Visual counts of rickettsial growth at lower concentrations of the antibiotics (0.25 to 0.0078 microgram/ml) showed that the Karp strain was 16 times more susceptible that the AFSC-4 strain to doxycycline. Azithromycin was much more effective than doxycycline against AFSC-4, inhibiting rickettsial growth at 0.0156 microgram/ml to levels below that achieved by 0.25 microgram of doxycycline per ml. Azithromycin was also more effective than doxycycline against the Karp strain, causing greater reductions in the number of rickettsiae per cell at lower concentrations. If in vivo testing confirms the in vitro effectiveness of azithromycin, it may prove to be the drug of choice for the treatment of scrub typhus in children and pregnant women, who should not take doxycycline, and in patients with refractory disease from locations where doxycycline-resistant strains of R. tsutsugamushi have been found. When tested in an in vitro assay system, azithromycin was more effective than doxycycline against doxycycline-susceptible and -resistant strains of R. tsutsugamushi.

Animals↗
PubMed1970

Plaque assay for Q fever and scrub typhus rickettsiae.

The plaque assay procedure developed for spotted fever and typhus group rickettsiae is also appropriate for scrub typhus and Q fever rickettsiae. The plaque titers of suspensions of Rickettsia tsutsugamushi and Coxiella burnetii compared favorably with end points obtained by titrations in mice.

Animals↗