斑点热群立克次体新种──虎林－93株的分离和鉴定

本文用鸡胚卵黄囊培养法及实验动物感染法从黑龙江省虎林县境内采集的嗜群血蜱中分离到了一株立克次体。命名为ＨＬ－９３株立克次体。经形态学及血清学试验证实为斑点热群立克次体。分别用微量免疫荧光法、十二烷基硫酸钠－聚丙烯酰胺凝胶不连续电泳法、单克隆抗体免疫酶斑点法、单克隆和多克隆抗体免疫印迹法、聚合酶链反应和限制性片段长度多态性分析法对该分离株进行鉴定并同已知国际标准株、国际及国内参考株进行比较。结果表明;ＨＬ－９３株立克次体无论在抗原性多肽上,还是在基因水平上在斑点热群立克次体中都是独特的。根据目前立克次体分类法,ＨＬ－９３株立克次体可以考虑是斑点热群立克次体的新种。     

Rickettsia serosurvey in Kimberley, Western Australia.

To determine if antibodies to rickettsiae (scrub typhus, spotted fever, and typhus group rickettsiae) occur among persons living in the Kimberley (northern tropical) region of Western Australia, 920 sera collected in a non-random manner in 1996 from patients in Kununurra, Broome, Fitzroy Crossing, Wyndham, Derby, and Halls Creek were tested by micro-immunofluorescence for antibodies to a panel of rickettsial antigens. Of 920 sera examined, 52 (5.6%) were positive for antibodies to one or more of the three groups of rickettsial microorganisms. The largest group of sera (24; 2.6%) were positive for scrub typhus (Orientia tsutsugamushi). Eleven other sera (1.2%) were positive for scrub typhus and spotted fever group rickettsiae and four (0.4%) were positive for scrub typhus, spotted fever group, and typhus group rickettsiae. In addition 13 sera (1.4%) were positive only for spotted fever group rickettsiae. In this study, only titers > or = 1:256 were considered significant. Thus, there is serologic evidence for scrub typhus and spotted fever group rickettsial infections in the Kimberley region of Western Australia. Because of the method of serum collection, it is not possible to determine the prevalence of seropositivity, but the data support the need for a proper epidemiologic study of rickettsial diseases in this region of Australia.     

Rickettsial antibody prevalence in southern Israel: IgG antibodies to Coxiella burnetii, Rickettsia typhi, and spotted fever group rickettsiae among urban- and rural-dwelling and Bedouin women

A retrospective serological survey was carried out using sera obtained from women at childbirth in the southern desert region of Israel to determine exposure experience to three rickettsial agents: Coxiella burnetii, Rickettsia typhi, and spotted fever group rickettsiae. Using the indirect fluorescent antibody method for determining IgG antibodies, it was found that about 40% of all sera examined demonstrated antibodies to one or more rickettsiae. Bedouin women appeared to be at greater risk of having antibodies to C. burnetii and spotted fever group rickettsiae than did Jewish residents of Beersheba, agricultural settlements, and development towns. The residents of development towns appeared to be at lower risk of developing antibodies to spotted fever group rickettsiae than did other populations sampled. Possible reasons for these differences are discussed.     

Rickettsia rickettsii and Rickettsia montana from Ixodid ticks in Connecticut

Dermacentor variabilis, infected with spotted fever group rickettsiae, parasitized 8 of 70 raccoons captured in Newtown, Connecticut. The spotted fever agent, Rickettsia rickettsii, was isolated and identified from 4 adult D. variabilis and from 1 nymphal Ixodes texanus removed from raccoons. This verifies the presence of this etiologic agent in ticks in an area where 6 people had clinical signs and symptoms of Rocky Mountain spotted fever (RMSF) and antibodies to R. rickettsii. These are the first isolations of R. rickettsii from D. variabilis in southern New England and the first identified rickettsiae from I. texanus. No rickettsiae were isolated from Ixodes muris or I. cookei. Rickettsia montana was recovered in Vero cell culture from a D. variabilis collected in East Haddam, Connecticut where RMSF is not known to be prevalent.     

Rickettsial and serologic evidence for prevalent spotted fever rickettsiosis in inner Mongolia

A field study in northeastern Inner Mongolia, People's Republic of China, in June of 1985 demonstrated a spotted fever group rickettsiosis. Two strains of spotted fever group rickettsiae were isolated. One strain was obtained from the blood of a patient with an eschar, regional lymphadenopathy, and history of a recent tick bite. The other strain originated from ova of ticks, Dermacentor nuttalli. These represent the second isolate of a spotted fever group rickettsia from a human and the first isolate from tick ova in the People's Republic of China. Antibodies to these rickettsiae were demonstrated by indirect immunofluorescence in the sera of 4% of healthy children and 34% of healthy human adults, in 58% of sheep, and in 76% of cattle in the same location. Hemolymph test revealed rickettsiae in 6 of 36 D. nuttalli examined.     

Distribution and prevalence of spotted fever group rickettsiae in ticks from South Carolina, with an epidemiological survey of persons bitten by infected ticks.

Live ticks removed from human hosts were tested for the presence of spotted fever group rickettsiae from 1974 through 1976. Spotted fever group rickettsiae were detected in Dermacentor variabilis, Amblyomma americanum, Amblyomma maculatum, and Ixodes scapularis. Persons from whom positive ticks were removed in 1974 and 1975 were questioned about ensuing illness. Eleven of 51 persons from whom rickettsia-positive D. variabilis were removed reported some unexplained combination of symptoms compatible with Rocky Mountain spotted fever. Persons with positive ticks other than D. variabilis reported no illness in the 2 weeks following exposure. D. variabilis from the Coastal Plain of South Carolina have a higher percentage of infection than those from the Piedmont region. Conversely, most reported human cases of spotted fever are from the Piedmont.     

Evidence of rickettsial spotted fever and ehrlichial infections in a subtropical territory of Jujuy, Argentina.

Between November 1993 and March 1994, a cluster 6 pediatric patients with acute febrile illnesses associated with rashes was identified in Jujuy Province, Argentina. Immunohistochemical staining of tissues confirmed spotted fever group rickettsial infection in a patient with fatal disease, and testing of serum of a patient convalescing from the illness by using an indirect immunofluorescence assay (IFA) demonstrated antibodies reactive with spotted fever group rickettsiae. A serosurvey was conducted among 16 households in proximity to the index case. Of 105 healthy subjects evaluated by IFA, 19 (18%) demonstrated antibodies reactive with rickettsiae or ehrlichiae: 4 had antibodies reactive with Rickettsia rickettsii, 15 with Ehrlichia chaffeensis, and 1 with R. typhi. Amblyomma cajennense, a known vector of R. rickettsii in South America, was collected from pets and horses in the area. These results are the first to document rickettsial spotted fever and ehrlichial infections in Argentina.     

A focus of dogs and Rickettsia massiliae-infected Rhipicephalus sanguineus in California

A recurrent focus of Rhipicephalus sanguineus infestation was investigated in a suburban area of southern California after reports of suspected Rocky Mountain spotted fever in two dogs on the same property. Abundant quantities of Rh. sanguineus were collected on the property and repeatedly from each dog, and Rickettsia massiliae DNA was detected by polymerase chain reaction (PCR). Whole blood and serum samples from four dogs were tested by using PCR and microimmunofluorescent assay for antibodies against spotted fever group rickettsiae. Serum samples from all four dogs contained antibodies reactive with R. massiliae, R. rhipicephali, R. rickettsii, and 364D Rickettsia but no rickettsial DNA was detected by PCR of blood samples. Serum cross-absorption and Western blot assays implicated R. massiliae as the most likely spotted fever group rickettsiae responsible for seropositivity. To our knowledge, this is the first detection of R. massiliae in ticks in California.     

Spotted fever group rickettsiosis in Chiba Prefecture]

In Chiba Prefecture, the first patient of infection with spotted fever group Rickettsia was found in 1987, thereafter nine patients were detected serologically by the end of 1990. Patients were found in the villages of Amatsukominato, Katsuura and Ootaki, which were located in southern part of Chiba Prefecture. The illness occurred from June to October. On the other hand, patients with Rickettsia tsutsugamushi were seen from October to September. Difference of prevalent seasons of these two types of rickettsiosis is important to make a clinical diagnosis and serological identification of this spotted fever group rickettsiosis. Antibody of the patients showed the highest titer to YH strain of Rickettsia and showed high cross-reactivity to other spotted fever group rickettsiae. For the diagnosis of the patient serologically, it was confirmed that any strain of spotted fever group Rickettsia were useful.     

Antigenic analysis of Chinese strains of spotted fever group rickettsiae by protein immunoblotting

Protein immunoblotting demonstrated that 6 Chinese strains of spotted fever group (SFG) rickettsiae from northern China had antigenic polypeptides identical to those of Rickettsia siberica (strains 246 and 232), and dissimilar to other SFG rickettsiae. The various species of other SFG rickettsiae exhibited serologically distinct epitopes as well as many cross-reactive epitopes in protein immunoblotting. All SFG rickettsiae examined in this study had major antigenic polypeptides of 113-160 kDa.     

Rocky Mountain spotted fever in Argentina

We describe the first molecular confirmation of Rickettsia rickettsii, the cause of Rocky Mountain spotted fever (RMSF), from a tick vector, Amblyomma cajennense, and from a cluster of fatal spotted fever cases in Argentina. Questing A. cajennense ticks were collected at or near sites of presumed or confirmed cases of spotted fever rickettsiosis in Jujuy Province and evaluated by polymerase chain reaction assays for spotted fever group rickettsiae. DNA of R. rickettsii was amplified from a pool of A. cajennense ticks and from tissues of one of four patients who died during 2003-2004 after illnesses characterized by high fever, severe headache, myalgias, and petechial rash. The diagnosis of spotted fever rickettsiosis was confirmed in the other patients by indirect immunofluorescence antibody and immunohistochemical staining techniques. These findings show the existence of RMSF in Argentina and emphasize the need for clinicians throughout the Americas to consider RMSF in patients with febrile rash illnesses.     

Serologic evidence of infection with ehrlichiae and spotted fever group rickettsiae among residents of Gag Island,Indonesia

The causative agents of scrub and murine typhus are considered endemic to Indonesia. However, the presence of spotted fever group rickettsiae and ehrlichiae have not been previously described in this country. During an investigation of arthropod-borne diseases on Gag Island, located northwest of the island of New Guinea in eastern Indonesia, the prevalence of antibody to the etiologic agents of monocytic ehrlichiosis, spotted fever rickettsiosis, and scrub and murine typhus were determined. Analysis of 55 blood samples from residents of Gag Island showed seroreactivity to antigen preparations of Ehrlichia chaffeensis (7 of 48, 14.6%), two spotted fever group rickettsiae: Rickettsia rickettsii (5 of 48, 10.4%) and R. conorii (10 of 49, 20.4%), Orientia tsutsugamushi (5 of 53, 9.4%), and R. typhi (1 of 48, 2.1% [by an indirect immunofluorescence assay] and 1 of 50, 2.0% [by an enzyme-linked immunosorbent assay]). These results show serologic evidence of infection with ehrlichiae and spotted fever group rickettsiae for the first time in Indonesia in a location where the prevalence of antibody to O. tsutsugamushi and R. typhi was lower.     

Ixodes ricinus: vector of a hitherto undescribed spotted fever group agent in Switzerland.

A tick/rickettsial survey in various parts of Switzerland revealed the presence of a new, hitherto undescribed spotted fever group rickettsia ("Swiss agent") in up to 11.7% of I. ricinus collected off vegetation. Infection in ticks was found to be generalized with rickettsiae developing intracellularly and occasionally also intranuclearly. As a result of massive growth in ovarial tissues, including the germinative cells, the rate of transovarial and filial infection was 100%. The "Swiss agent" appears to be nonpathogenic for guinea pigs, domestic rabbits, and Swiss mice, but in male meadow voles (Microtus pennsylvanicus) it produces a microscopically detectable infection in the tunica vaginalis. The rickettsia grows well in tissue culture systems including chick embryo fibroblast, Vero, and vole tissue cells, when inoculated via yolk sac into 5-day-old hens' eggs, it kills 100% of the embryos after 5 to 7 days. Antigenic relatedness of the "Swiss agent" to rickettsiae of the spotted fever group was indicated by indirect and direct fluorescent antibody staining. Preliminary serologic typing by microimmunofluorescence and by microagglutination indicated that the "Swiss agent" differs from all prototype strains of spotted fever group rickettsiae studied so far.     

Spotted fever group rickettsiosis on Awaji Island, Japan

Three cases of spotted fever group rickettsiosis occurring on Awaji Island, Hyogo Prefecture, the first reported in the Kinki District, are described. The illness appeared from August to September in 1988. High-grade fever and papular erythema were observed in all cases. Eschar, lymphadenopathy and hepatomegaly were observed in two-thirds of the cases, respectively. The antibody titers for Proteus OX-2 by the Weil-Felix reaction were elevated, and immunofluorescence test with R. rickettsii of the spotted fever group demonstrated a significant rise in specific antibody titer. Tetracycline was effective in all cases. Every patient was thought to have been infected with rickettsiae in the Yuzuruha mountain range in the southern part of Awaji Island. On the other hand, tsutsugamushi disease occurs in autumn and winter in the northern part of Awaji Island. It would be of considerable interest from an epidemiological viewpoint to study the vector of these spotted fever group rickettsiae.     

A highly sensitive and specific real-time PCR assay for the detection of spotted fever and typhus group Rickettsiae

A highly specific real-time polymerase chain reaction (PCR) assay was developed to detect spotted fever and typhus group rickettsiae using the citrate synthase gene as the target. The assay amplified rickettsial members of the spotted fever and typhus group including Rickettsia akari, R. australis, R. conorii, R. honei, "R. marmionii," R. sibirica, R. rickettsii, R. typhi, and R. prowazekii. The ancestral group rickettsia, R. bellii, did not produce a positive reaction, nor did other members of the order Rickettsiales or any non-rickettsial bacteria. The assay had a sensitivity of one target copy number per reaction as determined by serial dilutions of a plasmid containing a spotted fever group target sequence. This quantitative assay is useful for the enumeration of rickettsiae in clinical specimens and the diagnosis of rickettsial illnesses, when rickettsial numbers are very low.     

广东连平县存在新的蜱传斑点热立克次体

目的对采自广东省连平县的3种硬蜱进行蜱传斑点热立克次体检测分析。方法斑点热立克次体的ompA基因片段扩增并测定扩增片段的DNA序列。结果20只蜱分为17组,其中15组PCR检测阳性。对4个测序成功的标本进行聚类分析,证实其中3个序列单独聚为一类,与引起Flinders岛蜱传斑点热的弗诺立克次体(R.honei)、非洲蜱咬热的非洲立克次体(R.africa)、未定名斑点热的斯洛伐克立克次体(R.slovaca)以及西伯利亚立克次体BJ-90株等亲缘关系较近,另一序列与我国长白山地区检测到的JL-02具有较高的同源性。结论本研究提示该地区除了已证实的西伯利亚斑点热外,还存在新的蜱传斑点热。     

T cells mediate cross-protective immunity between spotted fever group rickettsiae and typhus group rickettsiae

Rickettsioses are severe infections caused by obligately intracellular bacteria that preferentially infect the endothelium lining the vasculature. The causative agents, rickettsiae, have been divided according to biological, genetic, and antigenic parameters into 2 main groups: spotted fever and typhus. They have not been thought to stimulate cross-reactive protective immune responses; however, in this study, we show that, in relevant animal models that mimic human rickettsial infections, there is reciprocal immunological cross-protection between spotted fever group and typhus group rickettsiae. Furthermore, we present evidence that T cells are responsible for this cross-immunity and that cross-stimulation of T cells also occurs in humans.     

Prevalence of spotted fever rickettsial antibodies in dogs and rodents in the Philippines

Antibodies against spotted fever group rickettsiae have been detected in blood samples of dogs and rodents obtained from selected areas in the Philippines. In this serosurvey, the positive percentage rates are 8.3% (11/132) in dogs and 12.2% (6/49) in rats. Positive results were read from samples tested with Rickettsia japonica antigen. No positive result was obtained in blood samples of rats and house mice using R. akari antigen. The findings of this study are the first to confirm the detection of spotted fever group rickettsial antibodies in the Philippines.     

Spotted fever group Rickettsia in Yunnan Province, China

Information about spotted fever group (SFG) rickettsiae in southern China remains sparse. A specific and sensitive real-time PCR assay for detection of SFG rickettsiae was established and used to detect the prevalence rate of SFG rickettsiae in Yunnan Province, China. The limit of detection (LOD) of our real-time PCR was 200 copies per reaction, which is more sensitive than the previously developed nested PCR assays for Rickettsia. We tested 265 blood samples (127 goats, 78 dogs, and 60 cattle) collected from Yunnan Province using the real-time PCR assay and revealed that the prevalence of SFG rickettsiae among dogs, cattle, and goats were 14.10%, 23.33%, and 24.41%, respectively. The SFG rickettsiae detected in animals in Yunnan Province were classified into two genotypes: a unique group that is different from all known SFG rickettsiae species, and R. heilongjiangensis.     

Aponomma hydrosauri, the reptile-associated tick reservoir of Rickettsia honei on Flinders Island, Australia

Rickettsia honei is the etiologic agent of Flinders Island (Australia) spotted fever. The tick Aponomma hydrosauri is associated with reptiles and is the arthropod reservoir for this rickettsia on Flinders Island. The rickettsia appears to be maintained in the tick via vertical transmission. Of 46 ticks examined, 29 (63%) were positive for spotted fever group rickettsiae by detection of the citrate synthase gene by a polymerase chain reaction (PCR). From the positive tick samples, seven were sequenced and found to be 100% homologous with R. honei. Of 17 reptiles examined, none had evidence of rickettsiae by PCR or culture of blood.