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Sensitive microplate enzyme-linked immunosorbent assay for detection of antibodies against the scrub typhus rickettsia, Rickettsia tsutsugamushi.

A microtiter enzyme-linked immunosorbent assay (ELISA) has been developed for the titration of antibodies against scrub typhus in human and animal sera. Scrub typhus rickettsiae were grown in monolayers of irradiated mouse LM3 cells and separated from host cell materials by differential centrifugation, filtration through a glass filter (AP-20, Millipore Corp.), and isopycnic banding in Renografin density gradients. The scrub typhus ELISA antigens were obtained from the purified viable rickettsiae by French pressure cell disruption and addition of 0.2% Formalin to the soluble extract. Antisera prepared in rabbits against the prototype Karp, the Kato, and the Gilliam strains of scrub typhus were used to standardize the ELISA and to compare its sensitivity and specificity to that of the indirect fluorescent antibody test (IFA). ELISA titers were measured as the greatest serum dilution showing an optical density 0.25 above controls or by the optical density achieved at a fixed serum dilution. The IFA and ELISA end point titers were quite similar, and all three measures of titer had comparable specificity for the strains of scrub typhus. No cross-reactions between the typhus and scrub typhus wera were observed by ELISA. Both the immunoglobulin M (IgM) and IgG antibody titers of 12 sequential sera from four patients with scrub typhus were obtained by IFA and ELISA. The IFA and ELISA end point titers for IgM and IgG had correlation coefficients of 0.91 and 0.97, respectively, whereas the ELISA optical density values at a serum dilution of 1:100 had slightly lower correlations with IFA titers (0.80 and 0.94). Early rising IgM titers followed by rising IgG titers were demonstrated by ELISA in three patients with primary scrub typhus infections, whereas the IgG response predominated in a patient with a reinfection. It is concluded that the ELISA for scrub typhus is a very satisfactory alternative to the IFA test.

Animals↗

Scrub typhus during pregnancy and its treatment: a case series and review of the literature.

Although scrub typhus is uncommon in pregnant women, when present, it can have serious repercussions for the mother and developing fetus. Since it is uncommon, the clinical impact of scrub typhus on pregnancy has not been elucidated and an effective and safe therapeutic regimen has not been validated. The medical records of pregnant women whose scrub typhus were treated at Chungnam National University Hospital were reviewed and their clinical outcomes were evaluated. A review of the literature was also performed on pregnant women with scrub typhus and their clinical outcomes. Eight pregnant women with scrub typhus were treated successfully with a single 500-mg dose of azithromycin, and no relapses were reported. They all delivered healthy babies at term, without congenital or neonatal complications. In the reviews, azithromycin was effective against scrub typhus and had favorable pregnancy outcomes. Ciprofloxacin and cefuroxime failed to treat scrub typhus and fetal loss resulted. A single 500-mg dose of azithromycin may be a reasonable treatment regimen for pregnant women with scrub typhus. Ciprofloxacin might not be advisable for the treatment of scrub typhus during pregnancy. Scrub typhus itself seems to have serious adverse effects on pregnancy if not appropriately controlled.

Adult↗

The natural history of recrudescent typhus (Brill-Zinsser disease) in Bosnia.

Between 1954 and 1963, typhus, both epidemic and recrudescent, has been studied in Bosnia. The recrudescent typhus cases occur sporadically throughout the year, whereas epidemic typhus occurs mainly between November and June. During 1954-63, some 10-20 years after the extensive typhus epidemics of the Second World War, the highest incidence of recrudescent typhus was in the 20-50 year age-group, whereas epidemic typhus reached its highest incidence in the 2-30 year age-group. Field observations support Zinsser's hypothesis that man is the interepidemic reservoir of typhus. Recrudescent typhus is not likely to cause epidemics of primary epidemic typhus in environments where lice are scarce. However, if typhus is to be eradicated, it seems that it will be necessary to destroy all lice in areas where there are people who have at some time had the disease.

Adolescent↗

HIV-1 suppression during acute scrub-typhus infection.

BACKGROUND: In HIV-1-infected individuals, viral load has been reported to rise transiently if an acute infection with another organism occurs. Our study was prompted by the unexpected finding that HIV-1 copy number fell during an acute infection with Orientia tsutsugamushi, the causative agent of scrub typhus. METHODS: Serial HIV-1 viral load determinations were made in ten Thai adults with scrub typhus, who were not receiving antiretroviral therapy, and in five HIV-1-infected patients who had other infections (four malaria, one leptospirosis), during and after acute infections. Sera from HIV-1-infected patients with scrub typhus and from mice immunised with O. tsutsugamushi were examined for HIV-1-suppressive activity. FINDINGS: Median viral load 3 days after admission was significantly lower in the scrub-typhus group than in patients with other infections (193% vs 376% of day 28 values, p=0.03). In four O. tsutsugamushi-infected patients HIV-1 RNA copy number fell by three-fold or more compared with day 28 values, and HIV-1 copy numbers were below the assay threshold in two patients with scrub typhus. Five of seven HIV-1 isolates from non-typhus patients with CD4 lymphocytes less than 200 cells/microL were syncytia-inducing variants, whereas all ten isolates from O. tsutsugamushi-infected individuals matched by CD4-cell count were non-syncytia inducing (p=0.03). Sera from an HIV-1-negative patient with scrub typhus had potent HIV-1-suppressive activity in vitro. Sera from typhus-infected mice inhibited HIV-1 syncytia formation and bound by immunofluorescence to HIV-1-infected lymphocytes. INTERPRETATION: HIV-1-suppressive factors are produced during some scrub-typhus infection and should be investigated further in the search for novel strategies for the treatment and prevention of AIDS.

Acute Disease↗

Gastrointestinal manifestations of septic patients with scrub typhus in Maharat Nakhon Ratchasima Hospital.

Scrub typhus is an acute febrile illness caused by Orientia induced vasculitis, which is common in Asia and the Pacific Islands and is sometimes also encountered in Western countries. Even though it can cause multi-organ dysfunctions, there is limited information regarding the relationship between scrub typhus infection and gastrointestinal dysfunction. Therefore, a cross-sectional study was conducted to discover the gastrointestinal manifestations of septic patients with scrub typhus infection. During the study period, 80 septic cases were recruited, and according to the results of immunofluorescent antibody testing (IFA), 20 (25%) were found to have scrub typhus infection. The most common gastrointestinal symptoms of scrub typhus patients were vomiting 13 (65%), nausea 12 (60%), diarrhea 9 (45%), and hametamesis or melena 5 (25%). Gastrointestinal signs included hepatomegaly 8 (40%), jaundice 7 (35%), and abdominal pain 4 (20%). Elevation of SGOT, SGPT, and alkaline phosphatase were 16 (80%), 14 (70%), and 16 (80%), respectively. Direct bilirubin was elevated in 19 (95%) of the cases and half of the cases had a low serum protein level. Of scrub typhus cases, 8 (40%) had eschars. The sites of eschars were mostly in hidden areas, such as on the back, genitalia and abdomen. Three of the five patients with eschar had hepatomegaly on ultrasound examination. The significant findings of the scrub typhus septic patients with eschar on endoscopic examination were gastritis in two cases, gastritis with gastric erosion in two cases, and one case showed a duodenal ulcer and erosion. The differentiating point for endoscopic findings in scrub typhus compared to the other causes was that the stomach lesions were more frequent and severe than the duodenal lesions. According to our endoscopic findings, physicians should be aware of gastric and duodenal lesions in febrile patients with gastrointestinal symptoms, such as abdominal pain or discomfort and indigestion. Scrub typhus can cause gastrointestinal and liver dysfunction.

Adult↗

Murine and epidemic typhus rickettsiae: how close is their relationship?

Typhus fever has occurred globally as epidemic and endemic disorders. In 1910, Brill reported a typhus-like illness which Zinsser and others determined to be recurrent epidemic typhus fever. Maxcy, in 1926, proposed rodents and fleas as reservoir and vector, respectively, of endemic typhus, which Dyer confirmed in 1930. Animals experimentally infected with epidemic typhus (Rickettsia prowazeki) are immune to murine typhus (Rickettsia typhi) and vice versa. Similar solid cross-immunity exists for humans. The two diseases are clinically similar in pathologic and serologic reactions. Human epidemic typhus presumably involved a man-louse-man cycle without an animal reservoir. This concept is now questioned. Antibodies to R. prowazeki have been reported in livestock in Africa, rats in Manila, and from flying squirrels and humans in the United States. R. prowazeki was recovered from blood specimens of goats, sheep, from ixodid ticks, louse, and flea-ectoparasites of flying squirrels, and tissues of flying squirrels. More than 20 cases of squirrel-related acute epidemic typhus have been reported in the United States. R. prowazeki has not been recovered from human cases. Chemical studies of R. prowazeki and R. typhi show genetic similarities but differences in genome size and degree of hybridization suggest that interconversions between the two agents do not occur rapidly in nature. It is proposed that, with time, their relatedness will become even closer.

Animals↗

Epidemiologic, clinical and laboratory features of scrub typhus in thirty Thai children.

BACKGROUND: Scrub typhus, a potentially fatal rickettsial infection, is common in Asia. Although serologic surveys suggested that as many as one-fourth of cases of scrub typhus might be in children, very few reports of childhood scrub typhus are available in the medical literature. OBJECTIVES: To document the clinical, laboratory and epidemiologic characteristics of pediatric patients with scrub typhus. METHODS: From January 1, 2000 to December 31, 2001, all pediatric patients at Chiang Mai University Hospital who had obscure fever for >5 days were tested for indirect immunofluorescent antibody (IFA) against Orientia tsutsugamushi, the causative organism of scrub typhus. Scrub typhus was diagnosed on the basis of either a single IFA titer against O. tsutsugamushi > or =1/400 or a 4-fold or greater rise in IFA titer to at least 1/200. RESULTS: Thirty children with scrub typhus were enrolled. Most were diagnosed during the rainy months of June to November. Common physical signs included lymphadenopathy (93%), hepatomegaly (73%), eschar (68%), conjunctival hyperemia (33%), maculopapular rash (30%) and splenomegaly (23%). Eleven patients had interstitial pneumonitis and 1 patient had meningitis. All patients responded well to doxycycline or chloramphenicol. The average interval to defervescence after treatment was 29 h (range, 6 to 72). CONCLUSIONS: Clinical and epidemiologic features of 30 pediatric patients with scrub typhus are reported in a prospective study. The presence of eschar was helpful in making the diagnosis. Complications included pneumonitis and meningitis. All cases responded well to treatment with antibiotic.

Adolescent↗

[Epidemiology of scrub typhus in Matsu Peikang Island].

A retrospective survey of the epidemiology of scrub typhus in 1990 in Matsu Peikang Island was made at the end of December of 1990. The diagnosis of scrub typhus was based on the presence of escar, typical clinical manifestation and good response to tetracycline treatment. There were 89 cases of scrub typhus studied, and except for one civilian, the other 88 cases were military personnel, occurring in 2.2% of the total military personnel on the island. However it occurred in 0.08% of the civilian population. Seasonal distribution of scrub typhus was limited to between June and October of 1990. The peak incidence occurred in July. Scrub typhus was distributed over the whole island. A higher incidence of scrub typhus occurred in soldiers and sergeants than in officers. A significant different incidence was found in different units. No fatalities were reported. Different units. No fatalities were reported. Different military activities, duties and the location of the military unit may affect the incidence of scrub typhus. The scrub typhus epidemic was also related to temperature.

China↗

Outbreak of epidemic typhus associated with trench fever in Burundi.

BACKGROUND: After a 12-year absence, epidemic typhus has re-emerged among the displaced population of Burundi. Following the outbreak of civil war in 1993, over 760000 people now inhabit refugee camps, under appalling conditions. A typhus outbreak occurred among prisoners in a jail in N'Gozi in 1995. At the time, the disease was not recognised, and was referred to as sutama. Reports of sutama among the civilian population date back to late 1995 and, in association with body-louse infestation, the disease has subsequently swept across the higher and colder regions of the country. METHODS: During a field study in February, 1997, 102 refugees with sutama underwent clinical examination and interview. Serum samples were collected and infesting body lice removed. Microbiological analysis included antibody estimations and specific PCRs aimed at diagnosis of Rickettsia prowezekii, Bartonella quintana, and Borrelia recurrentis. Between January and September, 1997, nationwide epidemiological data on the prevalence and distribution of sutama was obtained through liaison with local health services. A second field study in March, 1997, entailed the collection of further serum samples from suspected cases of sutama in different regions of Burundi. FINDINGS: Most of the 102 patients with sutama during initial assessment presented with manifestations similar to those previously described for typhus in Africa, though skin eruptions occurred in only 25 (25%) cases. Microbiological testing revealed evidence of R prowazeki infection in 76 (75%) patients, confirming that most cases of clinically-diagnosed sutama were epidemic typhus, and supporting the reliability of clinical diagnosis as a basis for the nationwide surveillance of the disease. Up to September, 1997, 45558 typhus cases were clinically diagnosed, most of which occurred in regions at an altitude of over 1500 m. Serological testing of 232 individuals from different regions of Burundi provided microbiological evidence to support clinical diagnoses in seven provinces, confirming the widespread nature of the outbreak. Serum from 13 of the original 102 patients and 19 (8%) of the 232 suspected cases had raised antibody titres against B quintana. A fatality rate of 15% among jail inmates fell to 0.5% after administration of a single dose of 200 mg doxycycline to suspected cases. INTERPRETATION: A gigantic outbreak of R prowazekii-induced typhus and B quintana-induced trench fever is continuing in Burundi. Transmission of both diseases to such a large number of people has followed a widespread epidemic of body-louse infestation. Diagnosis of typhus could be reliably made by means of clinical criteria, and the disease could be efficiently and easily treated by antibiotics. This epidemic highlights the appalling conditions in central-African refugee camps and the failure of public-health programmes to serve their inhabitants. Louse-associated disease remains a major health threat in this and other war-torn regions of the world.

Anti-Bacterial Agents↗

Serosurvey of wild rodents for Rickettsioses (spotted fever, murine typhus and Q fever) in Java Island, Indonesia.

The prevalence of antibodies against spotted fever group rickettsia (SFGR), murine typhus and Q fever were investigated in wild rats captured in Indonesia. Sera of 327 rats were collected from Jakarta and Boyolali on Java Island. The prevalences of antibodies against SFGR and murine typhus were 128 (39.1%) and 48 (14.7%), respectively. Antibodies against Q fever were not detected in these serum samples. Antibodies against SFGR were found in all species of rats (20.8-51.9%). The antibody positive rate against murine typhus in Rattus norvegicus (38.0%) was significantly higher than that in other rat species (0-4.8%, p<0.01). The antibody positive rates against SFGR and murine typhus in rats captured in Jakarta were significantly higher than those in rats captured in Boyolali (p<0.01). In this survey, all species of rats had antibodies against SFGR, indicating that the 4 species of tested rats (R. norvegicus, R. rattus, R. exulans, R. tiomanicus) were infected with SFGR and that SFGR may infest the whole of Java Island. Most of the rats that were antibody-positive against murine typhus were captured in Jakarta. Therefore, R. norvegicus and R. rattus are likely to be important hosts of murine typhus in Jakarta. The antibody-positive rates against SFGR and murine typhus in rats captured in the dry season were significantly higher than those in rats captured in the rainy season. This may coincide with the active periods of ticks and fleas in Indonesia.

Animals↗

Murine typhus is a common cause of febrile illness in Bedouin children in Israel.

Murine typhus is known to be endemic among populations living in poverty and exposed to rats and their fleas. A prospective 2-y study was conducted to determine the contribution of murine typhus to undifferentiated febrile illnesses among Bedouin children attending an outpatient clinic in southern Israel. Children with fever > or = 38.5 degrees C lasting for > or = 3 d were enrolled in the study. Murine typhus was serologically confirmed by the microimmunofluorescence test. A total of 549 children met the inclusion criteria of whom 76 (13.8%) had serologically-confirmed murine typhus. The disease was diagnosed in 27 of 434 (6.2%) patients aged < 5 y and in 49 of 115 (42.6%) older children (p<0.001). Murine typhus was diagnosed in 54 of 288 (18.8%) patients between June and November and in 22 of 261 (8.4%) between December and May (p<0.001). Patients with murine typhus had significantly higher prevalence of anaemia, leukopenia, hyponatraemia, and elevated liver enzymes compared to children without the disease. A single child was hospitalized and all 76 patients recovered without complications. Murine typhus is an important cause of febrile illnesses among Bedouin children > or = 5 y of age living in southern Israel and usually runs a benign clinical course.

Animals↗

The clinical significance of upper gastrointestinal endoscopy in gastrointestinal vasculitis related to scrub typhus.

BACKGROUND AND STUDY AIMS: Scrub typhus is an acute febrile illness caused by Rickettsia tsutsugamushi-induced vasculitis, which is common in Asia and the Pacific islands and is sometimes encountered in Western countries because of increased travel and economic changes spurred by world globalization. Skin rash and eschar are typical physical findings on the trunk and extremities, but endoscopic mucosal changes have not been described in the gastrointestinal tract until now. We aimed to describe different endoscopic characteristics of the gastrointestinal manifestation of scrub typhus, to ascertain the necessity for endoscopy, and to determine correlations between the degrees of endoscopic lesion and clinical severity, including cutaneous manifestation. PATIENTS AND METHODS: Between January 1993 and October 1998, out of 256 scrub typhus patients, we applied esophagogastroduodenoscopy to 58 patients who complained of gastrointestinal symptoms but had no past history of these symptoms. We categorized clinical severity into four grades according to the degree of six clinical indicators of systemic complications, and endoscopic findings were graded from I to IV (I, normal, nonspecific hyperemia; II, distinct hyperemia, petechiae, purpura; II, superficial hemorrhage, erosion; IV, ulcer, active bleeding). RESULTS: Endoscopic findings of scrub typhus were characterized by petechiae, superficial hemorrhage, erosion, ulcers, and vascular bleeding (grade I, 14 patients; grade II, 11 patients; grade III, 16 patients; grade IV, 17 patients). In 83.3% of patients there was multiple occurrence of lesions without any predilection sites. Clinical severity was graded (grade I, 7 patients; grade II, 23 patients; grade III, 22 patients; grade IV, 6 patients). There was a correlation between clinical severity and endoscopic findings (P < 0.01). The grade of lesion was high in patients with cutaneous lesions (r(s) 0.359, P < 0.01). In two cases of gastric vascular bleeding, complete hemostasis was achieved by endoscopic hemoclipping. CONCLUSIONS: The major endoscopic features that can develop in scrub typhus are superficial mucosal hemorrhage, multiple erosions and ulcers without any predilection sites, and unusual vascular bleeding. The endoscopic features are related to cutaneous lesions and severity of the disease. Endoscopy is useful for diagnosis and management of gastrointestinal vasculitis related to scrub typhus.

Endoscopy, Gastrointestinal↗

Murine typhus in South Texas children.

BACKGROUND: Murine typhus is a zoonotic infection caused by Rickettsia typhi. This illness used to be endemic in the southeastern and gulf coast of the United States and is now only rarely reported in South Texas and Southern California. Murine typhus causes a febrile illness with headache and rash that has been well-described in adults. OBJECTIVE: To define the epidemiologic and clinical characteristics, laboratory findings, hospital course and response to therapy of children discharged from our hospital with a diagnosis of murine typhus. METHODS: Retrospective chart review of all children discharged from Driscoll Children's Hospital, Corpus Christi, TX, from January 1, 1990, to June 30, 1998, with a diagnosis of murine typhus. Patients. Thirty children (17 females) ages 2 to 17 years (mean, 10 +/- 4 years). RESULTS: Eighty percent of the children were admitted between May and November, and 67% had a history of contact with or exposure to a potential animal reservoir. Children were admitted after a mean of 7 +/- 4 days. The most common clinical features were fever 100%, rash 80% and headache 77%. Laboratory abnormalities included elevated erythrocyte sedimentation rate (75%), elevated serum transaminases (67%), hyponatremia (66%) and increased immature leukocytes without leukocytosis (63%). Only one child had leukocytosis and 40% had leukopenia. Defervescence occurred a mean of 35 +/- 19 h after initiation of appropriate antibiotics. Hospitalization lasted for a mean of 7 +/- 3 days. There were no readmissions and no patients died as a result of the infection. CONCLUSIONS: Despite being rarely reported in this country now, murine typhus continues to be an important cause of fever and hospitalization for children in South Texas. Children with murine typhus develop an illness similar to that reported in adults with fever, rash and headache. Children respond quickly to therapy with doxycycline or tetracycline and recover completely from their illness.

Adolescent↗

Prevalence of vectors of the spotted fever group Rickettsiae and murine typhus in a Bedouin town in Israel.

A survey of the vectors of spotted fever group Rickettsiae and of murine typhus was carried out in Rahat, a Bedouin town in the Negev Desert, where the diseases are endemic. Houses with known cases of spotted fever group Rickettsiae or murine typhus were compared with those without reported clinical cases. A neighboring Jewish community, Lehavim, where no cases of spotted fever group Rickettsiae and murine typhus were reported in recent years, was used as a control. In the houses of patients with spotted fever group Rickettsiae in Rahat, an average of 7.4 times more ticks were found than in control houses. Out of 190 ticks isolated from sheep and goats or caught by flagging in Rahat, 90% were Rhipicephalus sanguineus (Latreille), 7.9% Rhipicephalus turanicus Pomerantzev, and 2.1% were Hyalomma sp. In the houses of patients with murine typhus, three times more rats were caught and, on the average, each rat was infested with 2.2 times more fleas than rats in the control houses. Out of 323 fleas collected from 35 Norwegian rats (Rattus norvegicus Berkenhout), 191 were Xenopsylla cheopis Rothschild and 132 Echidnophaga murina Tiraboschi. Thus, there was a six to seven times higher probability of encountering a tick or flea vector where infections had occurred than in control houses in Rahat. The percentage of rats seropositive to Rickettsia typhi was similar in study and control households (78.3 and 76.2, respectively). In the control settlement, Lehavim, only three Mus musculus L. were caught, which were not infested with ectoparasites and their sera were negative for murine typhus. Out of 10 dogs examined in this settlement, 15 R. sanguineus and eight specimens of the cat flea (Ctenocephalides felis felis Bouché) were isolated. No rats were caught in this settlement. These data indicate that there is a correlation among the density of domestic animals, their ectoparasites, and the incidence of spotted fever group Rickettsiae and murine typhus in Rahat.

Animals↗

Murine typhus identified as a major cause of febrile illness in a camp for displaced Khmers in Thailand.

Scrub and murine typhus have been identified as causes of illness among the 238,000 displaced Khmer people residing in temporary settlements on the Thai side of the Thai-Cambodian border. Still, the true extent of the problem and the relative frequency of infection with scrub typhus as compared to murine typhus are unknown. We evaluated consecutive patients with unexplained pyrexia (documented fever, no exclusionary diagnosis, and constitutional symptoms) in 1 temporary settlement over 1 month. Laboratory studies included culture of blood and assay of paired sera for rickettsial IgM and IgG antibody, for dengue IgM and IgG antibody, and for leptospiral IgM and IgG antibody. Among 37 patients (27 adults and 10 children), 28 (75%) had a rickettsiosis (26 cases of murine typhus and 2 cases of scrub typhus). No case of enteric fever, dengue, or leptospirosis was diagnosed. The illnesses of 9 patients were not identified. Signs and symptoms did not distinguish confirmed rickettsial infections from undiagnosed illnesses. The 1 month attack rate of rickettsial infection was 29/100,000 for children and 185/100,000 for adults. Murine typhus was a major cause of febrile illness in this settlement.

Adult↗