Australian Rickettsial Reference Laboratory

Geelong, Australia

Australian Rickettsial Reference Laboratory

Geelong, Australia
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PubMed | Solomon Islands National University, National Referral Hospital, London School of Hygiene and Tropical Medicine, Australian Rickettsial Reference Laboratory and 3 more.
Type: Journal Article | Journal: Western Pacific surveillance and response journal : WPSAR | Year: 2016

To identify the etiology and risk factors of undifferentiated fever in a cluster of patients in Western Province, Solomon Islands, May 2014.An outbreak investigation with a case control study was conducted. A case was defined as an inpatient in one hospital in Western Province, Solomon Islands with high fever (>38.5 C) and a negative malaria microscopy test admitted between 1 and 31 May 2014. Asymptomatic controls matched with the cases residentially were recruited in a ratio of 1:2. Serum samples from the subjects were tested for rickettsial infections using indirect micro-immunofluorescence assay.Nine cases met the outbreak case definition. All cases were male. An eschar was noted in five cases (55%), and one developed pneumonitis. We did not identify any environmental factors associated with illness. Serum samples of all five follow-up cases (100%) had strong-positive IgG responses to scrub typhus. All but one control (10%) had a moderate response against scrub typhus. Four controls had low levels of antibodies against spotted fever group rickettsia, and only one had a low-level response to typhus group rickettsia.This outbreak represents the first laboratory-confirmed outbreak of scrub typhus in the Western Province of Solomon Islands. The results suggest that rickettsial infections are more common than currently recognized as a cause of an acute febrile illness. A revised clinical case definition for rickettsial infections and treatment guidelines were developed and shared with provincial health staff for better surveillance and response to future outbreaks of a similar kind.

Graves S.R.,Australian Rickettsial Reference Laboratory | Graves S.R.,Pathology North John Hunter HospitalNSW | Islam A.,Australian Rickettsial Reference Laboratory | Islam A.,Pathology North John Hunter HospitalNSW
American Journal of Tropical Medicine and Hygiene | Year: 2016

Q fever is endemic in Australia, and during the period 2005-2013 our laboratory diagnosed 379 cases in New South Wales. To evaluate clinical symptoms, epidemiology, mode of diagnosis, antibody profiles, and treatment, a subset of 160 (42%) Q fever cases were analyzed in detail following the return of a questionnaire by the patient's doctor and from their laboratory reports. Overall, 82% patients were male and predominantly middle aged. The majority of patients (89%) had animal contact among which 63% were with cattle, 11% with sheep, and 7% with kangaroos. Clinical symptoms were nonspecific: myalgia (94%), fever (91%), headache (80%), acute fatigue (64%), and arthralgia (55%). Most cases (93%) were acute, and serology (immunofluorescence) was the main diagnostic modality. Positive real-time polymerase chain reaction results were useful in the diagnosis of both acute and chronic Q fever, as was the isolation of Coxiella burnetii in cell culture. Doxycycline was the antibiotic most commonly used. © Copyright 2016 by The American Society of Tropical Medicine and Hygiene.

Chalada M.J.,Central Queensland University | Stenos J.,Australian Rickettsial Reference Laboratory | Bradbury R.S.,Central Queensland University
One Health | Year: 2016

Lyme Borreliosis is a common tick-borne disease of the northern hemisphere caused by the spirochaetes of the Borrelia burgdorferi sensu lato (B. burgdorferi s. l.) complex. It results in multi-organ disease with arthritic, cardiac, neurological and dermatological manifestations. In the last twenty-five years there have been over 500 reports of an Australian Lyme-like syndrome in the scientific literature. However, the diagnoses of Lyme Borreliosis made in these cases have been primarily by clinical presentation and laboratory results of tentative reliability and the true cause of these illnesses remains unknown. A number of animals have been introduced to Australia that may act as B. burgdorferi s. l. reservoirs in Lyme-endemic countries, and there are some Australian Ixodes spp. and Haemaphysalis spp. ticks whose geographical distribution matches that of the Australian Lyme-like cases. Four published studies have searched for Borrelia in Australian ticks, with contradicting results. The cause of the potential Lyme-like disease in Australia remains to be defined. The evidence to date as to whether these illnesses are caused by a Borrelia species, another tick borne pathogen or are due to a novel or unrelated aetiology is summarised in this review. © 2016 The Authors.

Lockhart M.,Australian Rickettsial Reference Laboratory | Islam A.,NSW Health | Graves S.,NSW Health | Fenwick S.,Murdoch University | Stenos J.,Australian Rickettsial Reference Laboratory
FEMS Immunology and Medical Microbiology | Year: 2012

Coxiella burnetii is an acidophilic, intracellular bacterium that causes the human disease Q fever. In some studies, it is important to distinguish between viable and nonviable C. burnetii. We compared four methods for detecting and measuring viable C. burnetii in biological samples as follows: growth in two different cell culture lines, infection of severe combined immunodeficient (SCID) mice (leading to death) and infection of SCID mice with detection of C. burnetii in their spleen (after euthanasia at day 50 postinfection). Two isolates of C. burnetii were used ('Henzerling' and 'Arandale'). Our in-house qPCR assay for C. burnetii DNA was used as a control. SCID mouse inoculation was more sensitive than cell culture. The assay that detected C. burnetii in SCID mouse spleens was slightly more sensitive than SCID mice deaths alone. Approximately one viable C. burnetii cell could be detected by this method, making it suitable for determining the viability of C. burnetii in a sample. © 2011 Federation of European Microbiological Societies.

Sukocheva O.A.,Hanson Institute | Marmion B.P.,Hanson Institute | Storm P.A.,Hanson Institute | Lockhart M.,Australian Rickettsial Reference Laboratory | And 2 more authors.
QJM | Year: 2010

Background: Previous studies of inciting factors for a prolonged post-infection fatigue syndrome after Q fever (variously termed QFS or Q fever associated CFS/ME in the literature) showed that after the acute infection a high proportion of asymptomatic and QFS patients had Q fever antibody and also low levels in PBMC and bone marrow of Coxiella burnetii (C.b.) DNA with PCR assays directed against three different target sequences in different parts of the coxiella genome. Attempts to isolate a strain of C.b. in A/J mice, and cell culture from PCR positive PBMC and bone marrow were consistently negative. The detailed composition of the persisting coxiella residues remains to be defined. Aim: To retest and provide detailed results on selected PCR positive samples from the Birmingham Q fever outbreak patients tested by a highly sensitive method to detect viable organisms and to determine the nature of the residual coxiella cell components. Design: Laboratory case study. Methods: NOD/SCID mice were inoculated with samples from the 1989 Q fever outbreak in Birmingham and followed for evidence of infection and the presence of coxiella DNA and specific antigens in spleen and liver macrophages. A significant, unexpected finding of specific antigen was followed by assessment of its ability to provoke production of inflammatory and non-inflammatory cytokines in mice, in THP-1 human macrophage cell cultures and to induce inflammatory lesions in the skin of guinea pigs hyperimmunized against Q fever vaccine. Results: Culture of samples from 10 Birmingham Q fever patients in NOD/SCID mice, 12 years from infection did not yield viable Coxiella burnetii, as shown earlier. However complexes of material with coxiella antigens were found in mouse spleens in all cases but in significantly greater amounts in samples from those with post Q fever fatigue syndrome. The antigenic complexes [now designated 'immunomodulatory complexes' (IMC)] were shown to stimulate cytokine release in the mice and in the THP-1 macrophages and to provoke an inflammatory reaction on intradermal injection into the skin of Q fever hyperimmunized guinea pigs. Conclusion: The study identifies a non-infective complex of C.b. antigens able to survive in the host and provoke aberrant humoral and cell medicated immunity responses - a possible pathogenic link between initial infection and a subsequent long-term post Q fever fatigue syndrome. © The Author 2010. Published by Oxford University Press on behalf of the Association of Physicians. All rights reserved.

Islam A.,John Hunter Hospital | Islam A.,Australian Rickettsial Reference Laboratory | Ferguson J.,John Hunter Hospital | Givney R.,John Hunter Hospital | And 2 more authors.
American Journal of Tropical Medicine and Hygiene | Year: 2011

Exposure to Coxiella burnetii is a risk in the Hunter New England (HNE) region of New South Wales (NSW), Australia, based on yearly reported cases of Q fever. We assessed seroprevalence of phase II antibodies to C. burnetii by indirect immunofluorescence assay (IFA; screening at 1/50 dilution) of residents of 24 local government areas (LGA) of the HNE region of NSW. A total of 2,438 randomly selected sera sent to the Hunter Area Pathology Service for routine diagnostic purposes (not Q fever testing) during the period of 2006-2009 were tested. The overall seroprevalence in sample group was 7%. The proportion of males (59%) was higher than females (41%). In age distribution, the largest proportion (37%) of seropositives was in the > 60 years age group. Lower prevalence was observed in 0-9 years (1%) and 10-19 years (5%) age groups. The seroprevalence in different LGAs varied between 0.5% and 22%. It was highest in Guyra (22%), Gunnedah (21%), Tenterfield (18%), and Narrabri (16%), with Newcastle (0.5%), Port Stephens (2%), Lake Macquarie (3%), and Singleton (3%) being the lowest. In most of the LGAs, seroprevalence was between 6% and 12%. This report indicates a considerable exposure to C. burnetii of residents in rural areas of the HNE region and is consistent with the high notification rate for Q fever in this part of Australia. Copyright © 2011 by The American Society of Tropical Medicine and Hygiene.

Hii S.-F.,University of Queensland | Abdad M.Y.,Australian Rickettsial Reference Laboratory | Kopp S.R.,University of Queensland | Stenos J.,Australian Rickettsial Reference Laboratory | And 2 more authors.
Parasites and Vectors | Year: 2013

Background: The recent detection of Rickettsia felis DNA in dogs in Australia suggests that dogs are potential mammalian reservoir hosts for this emerging rickettsia. To date, there is no published report addressing the seroprevalence of R. felis in dogs in Australia. Methods. Antigens for R. felis were produced by inoculating confluent XTC-2 monolayer cell cultures with three pools of cat flea (Ctenocephalides felis) homogenates. Infection was confirmed by real-time (qPCR), conventional or nested PCRs targeting the ompB, gltA, 17 kDa and ompA genes. Two hundred and ninety-two dogs from Southeast Queensland and the Northern Territory were tested for the presence of R. felis antibodies using a microimmunofluorescence (IF) test and the seroprevalence and associated risk factors for exposure were determined using both uni- and multi-variate analyses. Results: Rickettsia felis was successfully isolated in cell culture from all three cat-flea pools. One hundred and forty-eight dogs (50.7%) showed seropositivity with titres ≥64 and 54 (18.5%) with titres ≥128. At antibody titres ≥64, dogs with active ectoparasite control were less likely to be seropositive to R. felis (OR: 2.60; 95% CI: 1.20 - 5.56). Conclusions: This first reported isolation of R. felis in cell culture in Australia allowed for the production of antigen for serological testing of dogs. Results of this serological testing reflects the ubiquitous exposure of dogs to R. felis and advocate for owner vigilance with regards to ectoparasite control on domestic pets. © 2013 Hii et al.; licensee BioMed Central Ltd.

Hii S.F.,University of Queensland | Kopp S.R.,University of Queensland | Abdad M.Y.,Australian Rickettsial Reference Laboratory | Thompson M.F.,University of Queensland | And 3 more authors.
Vector-Borne and Zoonotic Diseases | Year: 2011

Rickettsia felis causes flea-borne spotted fever in humans worldwide. The cat flea, Ctenocephalides felis, serves as vector and reservoir host for this disease agent. To determine the role of dogs as potential reservoir hosts for spotted fever group rickettsiae, we screened blood from 100 pound dogs in Southeast Queensland by using a highly sensitive genus-specific PCR. Nine of the pound dogs were positive for rickettsial DNA and subsequent molecular sequencing confirmed amplification of R. felis. A high prevalence of R. felis in dogs in our study suggests that dogs may act as an important reservoir host for R. felis and as a potential source of human rickettsial infection. © Copyright 2011, Mary Ann Liebert, Inc.

PubMed | University of Sydney, University of New South Wales and Australian Rickettsial Reference Laboratory
Type: Journal Article | Journal: QJM : monthly journal of the Association of Physicians | Year: 2016

A detailed description of the natural history of acute Q fever, caused by infection with Coxiella burnetii, AIM: : To significantly increase understanding of the illness.Subjects with provisional acute Q fever (n=115) were recruited from primary care in rural Australia, and followed prospectively by interview and blood collection including for serological confirmation. A nested series of subjects with prolonged illness (cases), and those without (controls), were investigated in detail.Total phase I and phase II anti-C. burnetii antibodies were detected by complement fixation test; and IgG, IgM and IgA phase I and phase II titres by immunofluorescence. Flow cytometric analysis was conducted to enumerate circulating T cells subsets, B cells, monocytes and natural killer cells.Serological testing confirmed acute Q fever in 73 subjects (63%). The acute illness featured fever, headache, sweats, fatigue and anorexia; and varied widely in severity, causing an average of 8 days in bed and 15 days out of work or other role in the first month of illness. The illness course varied from 2 days to greater than a year. No cases of chronic, localized Q fever infection, such as endocarditis, were identified. Neither severe nor prolonged illness were associated with persistence of C. burnetii DNA, altered patterns of C. burnetii-specific IgG, IgM or IgA antibody production, or altered leucocyte subsets.The severity of acute Q fever alone predicted prolonged duration. Further studies are warranted to better understand the pathophysiology of prolonged illness after acute Q fever.

PubMed | AgriBio Laboratory, University of Melbourne and Australian Rickettsial Reference Laboratory
Type: Journal Article | Journal: Clinical and vaccine immunology : CVI | Year: 2016

Although many studies have reported the indirect immunofluorescence assay (IFA) to be more sensitive in detection of antibodies to Coxiella burnetii than the complement fixation test (CFT), the diagnostic sensitivity (DSe) and diagnostic specificity (DSp) of the assay have not been previously established for use in ruminants. This study aimed to validate the IFA by describing the optimization, selection of cutoff titers, repeatability, and reliability as well as the DSe and DSp of the assay. Bayesian latent class analysis was used to estimate diagnostic specifications in comparison with the CFT and the enzyme-linked immunosorbent assay (ELISA). The optimal cutoff dilution for screening for IgG and IgM antibodies in goat serum using the IFA was estimated to be 1:160. The IFA had good repeatability (>96.9% for IgG, >78.0% for IgM), and there was almost perfect agreement (Cohens kappa > 0.80 for IgG) between the readings reported by two technicians for samples tested for IgG antibodies. The IFA had a higher DSe (94.8%; 95% confidence interval [CI], 80.3, 99.6) for the detection of IgG antibodies against C. burnetii than the ELISA (70.1%; 95% CI, 52.7, 91.0) and the CFT (29.8%; 95% CI, 17.0, 44.8). All three tests were highly specific for goat IgG antibodies. The IFA also had a higher DSe (88.8%; 95% CI, 58.2, 99.5) for detection of IgM antibodies than the ELISA (71.7%; 95% CI, 46.3, 92.8). These results underscore the better suitability of the IFA than of the CFT and ELISA for detection of IgG and IgM antibodies in goat serum and possibly in serum from other ruminants.

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