The immune response in a cat-related outbreak of Q fever as measured by the indirect immunofluorescence test and the enzyme-linked immunosorbent assay

1990 ◽  
Vol 36 (4) ◽  
pp. 292-296 ◽  
Author(s):  
J. Embil ◽  
J. C. Williams ◽  
T. J. Marrie
Keyword(s):  
Immune Response ◽  
Phase I ◽  
Phase Ii ◽  
Q Fever ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Whole Cells ◽  
Chronic Q Fever ◽  
Acute Q Fever ◽  
Indirect Immunofluorescent

The isotypic immune response of 16 individuals who developed Q fever pneumonia following exposure to an infected parturient cat was studied. The enzyme-linked immunosorbent (ELISA) test was used to detect IgM, IgA, and IgG antibodies to phase I and phase II Coxiella burnetii whole-cell antigens and to the phase I lipopolysaccharide. The indirect immunofluorescent antibody (IFA) test was also used to detect antibodies to phase I and phase II whole cells. None of the 16 subjects developed antibodies to the phase I lipopoly saccharide. The ELISA was more sensitive than the IFA test. IgM antibodies to phase II antigen were detectable by ELISA in 80% of the subjects at the time of onset of symptoms and were still present in 7 of the 8 tested at 32 weeks following the onset of symptoms. In all instances (ELISA: IgG, IgM; IFA: IgG, IgM) phase II antibodies developed earlier and reached higher levels than did phase I antibodies. The absence of antibodies to phase I lipopolysaccharide in acute Q fever combined with our unpublished findings of antibodies to phase I lipopoly saccharide in chronic Q fever suggests that this test may be used to distinguish acute from chronic Q fever. Key words: Q fever, immune response, ELISA.

scholarly journals Animals withCoxiella burnetiiInfection Demonstrate a Western Immunoblot Profile of Chronic Q Fever in Humans

1996 ◽  
Vol 7 (1) ◽  
pp. 45-48
Author(s):  
TJ Marrie ◽  
Linda Yates
Keyword(s):  
Immune Response ◽  
Phase I ◽  
Phase Ii ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Western Immunoblotting ◽  
Western Immunoblot ◽  
Chronic Q Fever

Western immunoblotting was used to compare the immune response toCoxiella burnetiiphase I and phase II antigens of humans with acute and chronic Q fever with that of infected cats, rabbits, cows and raccoons. The cats, rabbits, cows and raccoons had an immunoblot profile similar to that of the human with chronic Q fever.

scholarly journals Predominant Immunoglobulin A Response to Phase II Antigen of Coxiella burnetii in Acute Q Fever

1999 ◽  
Vol 6 (2) ◽  
pp. 173-177 ◽  
Author(s):  
Pierre-Edouard Fournier ◽  
Didier Raoult
Keyword(s):  
Phase I ◽  
Phase Ii ◽  
Rural Areas ◽  
Q Fever ◽  
Immunoglobulin A ◽  
Immunoglobulin M ◽  
Retrospective Case ◽  
Chronic Q Fever ◽  
Antibody Titers ◽  
Acute Q Fever

ABSTRACT Diagnosis of acute Q fever is usually confirmed by serology, on the basis of anti-phase II antigen immunoglobulin M (IgM) titers of ≥1:50 and IgG titers of ≥1:200. Phase I antibodies, especially IgG and IgA, are predominant in chronic forms of the disease. However, between January 1982 and June 1998, we observed anti-phase II antigen IgA titers of ≥1:200 as the sole or main antibody response in 10 of 1,034 (0.96%) patients with acute Q fever for whom information was available. In order to determine whether specific epidemiological or clinical factors were associated with these serological profiles, we conducted a retrospective case-control study that included completion of a standardized questionnaire, which was given to 40 matched controls who also suffered from acute Q fever. The mean age of patients with elevated phase II IgA titers was significantly higher than that usually observed for patients with acute Q fever (P = 0.026); the patients were also more likely than controls to live in rural areas (P = 0.026) and to have increased levels of transaminase in blood (P = 0.03). Elevated IgA titers are usually associated with chronic Q fever and are directed mainly at phase I antigens. Although the significance of our findings is unexplained, we herein emphasize the fact that IgA antibodies are not specific for chronic forms of Q fever and that they may occasionally be observed in patients with acute disease. Moreover, as such antibody profiles may not be determined by most laboratories, which test only for total antibody titers to phase I and II antigens, the three isotype-specific Ig titers should be determined as the first step in diagnosing Q fever.

scholarly journals Validation of a Novel Commercial ELISA Test for the Detection of Antibodies against Coxiella burnetii

Pathogens ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1075
Author(s):  
Salvatore Ledda ◽  
Cinzia Santucciu ◽  
Valentina Chisu ◽  
Giovanna Masala
Keyword(s):  
Diagnostic Accuracy ◽  
Phase Ii ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Animal Health ◽  
Elisa Test ◽  
Clinical Diagnostics ◽  
Complex Life Cycle ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specificity And Sensitivity

Q fever is a zoonosis caused by Coxiella burnetii, a Gram-negative pathogen with a complex life cycle and a high impact on public and animal health all over the world. The symptoms are indistinguishable from those belonging to other diseases, and the disease could be symptomless. For these reasons, reliable laboratory tests are essential for an accurate diagnosis. The aim of this study was to validate a novel enzyme-linked immunosorbent assay (ELISA) test, named the Chorus Q Fever Phase II IgG and IgM Kit (DIESSE, Diagnostica Senese S.p.A), which is performed by an instrument named Chorus, a new device in medical diagnostics. This diagnostic test is employed for the detection of antibodies against C. burnetii Phase II antigens in acute disease. Our validation protocol was performed according to the Italian Accreditation Body (ACCREDIA) (Regulation UNI CEI EN ISO/IEC 17025:2018 and 17043:2010), OIE (World Organization for Animal Health), and Statement for Reporting Studies of Diagnostic Accuracy (STARD). Operator performance was evaluated along with the analytical specificity and sensitivity (ASp and ASe) and diagnostic accuracy of the kit, with parameters such as diagnostic specificity and sensitivity (DSp and DSe) and positive and negative predictive values (PPV and NPV), in addition to the repeatability. According to the evaluated parameters, the diagnostic ELISA test was shown to be suitable for validation and commercialization as a screening method in human sera and a valid support for clinical diagnostics.

scholarly journals Questing one Brazilian query: reporting 16 cases of Q fever from Minas Gerais, Brazil

2006 ◽  
Vol 48 (1) ◽  
pp. 5-9 ◽  
Author(s):  
Paulo Sérgio Gonçalves da Costa ◽  
Marco Emilio Brigatte ◽  
Dirceu Bartolomeu Greco
Keyword(s):  
Phase I ◽  
Phase Ii ◽  
Fever Of Unknown Origin ◽  
Q Fever ◽  
Acute Infection ◽  
Case Series ◽  
Febrile Illness ◽  
Unknown Origin ◽  
Clinical Form ◽  
Chronic Q Fever

Q fever has been considered non-existing in Brazil where reports of clinical cases still cannot be found. This case-series of 16 patients is a result of a systematic search for such illness by means of clinical and serologic criteria. Serologic testing was performed by the indirect microimmunofluorescence technique using phase I/II C. burnetii antigens. Influenza-like syndrome was the most frequent clinical form (eight cases - 50%), followed by pneumonia, FUO (fever of unknown origin), mono-like syndrome (two cases - 12.5% each), lymphadenitis (one case - 6.3%) and spondylodiscitis associated with osteomyelitis (one case - 6.3%). The ages varied from four to 67 years old with a median of 43.5. All but one patient had positive serologic tests for phase II IgG whether or not associated with IgM positivity compatible with acute infection. One patient had both phase I and phase II IgG antibodies compatible with chronic Q fever. Seroconvertion was detected in 10 patients. Despite the known limitations of serologic diagnosis, the cases here reported should encourage Brazilian doctors to include Q fever as an indigenous cause of febrile illness.

scholarly journals Early Diagnosis and Treatment of Patients with Symptomatic Acute Q Fever Do Not Prohibit IgG Antibody Responses to Coxiella burnetii

2012 ◽  
Vol 19 (10) ◽  
pp. 1661-1666 ◽  
Author(s):  
C. C. H. Wielders ◽  
L. M. Kampschreur ◽  
P. M. Schneeberger ◽  
M. M. Jager ◽  
A. I. M. Hoepelman ◽  
...  
Keyword(s):  
Early Diagnosis ◽  
Phase I ◽  
Phase Ii ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Antibody Responses ◽  
Igg Antibody ◽  
Antibody Titers ◽  
Acute Q Fever

ABSTRACTLittle is known about the effect of timing of antibiotic treatment on development of IgG antibodies following acute Q fever. We studied IgG antibody responses in symptomatic patients diagnosed either before or during development of the serologic response toCoxiella burnetii. Between 15 and 31 May 2009, 186 patients presented with acute Q fever, of which 181 were included in this retrospective study: 91 early-diagnosed (ED) acute Q fever patients, defined as negative IgM phase II enzyme-linked immunosorbent assay (ELISA) and positive PCR, and 90 late-diagnosed (LD) acute Q fever patients, defined as positive/dubious IgM phase II ELISA and positive immunofluorescence assay (IFA). Follow-up serology at 3, 6, and 12 months was performed using IFA (IgG phase I and II). High IgG antibody titers were defined as IgG phase II titers of ≥1:1,024 together with IgG phase I titers of ≥1:256. At 12 months, 28.6% of ED patients and 19.5% of LD patients had high IgG antibody titers (P= 0.17). No statistically significant differences were found in frequencies of IgG phase I and IgG phase II antibody titers at all follow-up appointments for adequately and inadequately treated patients overall, as well as for ED and LD patients analyzed separately. Additionally, no significant difference was found in frequencies of high antibody titers and between early (treatment started within 7 days after seeking medical attention) and late timing of treatment. This study indicates that early diagnosis and antibiotic treatment of acute Q fever do not prohibit development of the IgG antibody response.

Solitary IgM phase II response has a limited predictive value in the diagnosis of acute Q fever

2012 ◽  
Vol 140 (11) ◽  
pp. 1950-1954 ◽  
Author(s):  
C. F. H. RAVEN ◽  
J. L. A. HAUTVAST ◽  
T. HERREMANS ◽  
A. C. A. P. LEENDERS ◽  
P. M. SCHNEEBERGER
Keyword(s):  
Phase Ii ◽  
Predictive Value ◽  
Q Fever ◽  
Immunofluorescence Assay ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Initial Sample ◽  
Acute Q Fever ◽  
Diagnostic Criterion

SUMMARYWe investigated the positive predictive value (PPV) of a solitary positive immunoglobulin M (IgM) phase II response for the serodiagnosis of acute Q fever detected with either an indirect immunofluorescence assay (IFA) or an enzyme-linked immunosorbent assay (ELISA). Initial and follow-up sera from patients suspected of acute Q fever were included if initially only IgM phase II tested positive with IFA in 2008 (n=92), or ELISA in 2009 (n=85). A seroconversion for Q fever was defined as an initial sample being IgG phase II negative but positive in the follow-up sample. The PPV of an initial isolated IgM phase II result detected by IFA or ELISA was 65% and 51%, respectively, and therefore appeared not to adequately predict acute Q fever. For this reason it cannot be used as a diagnostic criterion nor should it be included in public health notification without confirmation with other markers or a follow-up serum sample.

scholarly journals Real-Time PCR with Serum Samples Is Indispensable for Early Diagnosis of Acute Q Fever

2009 ◽  
Vol 17 (2) ◽  
pp. 286-290 ◽  
Author(s):  
Peter M. Schneeberger ◽  
Mirjam H. A. Hermans ◽  
Erik J. van Hannen ◽  
Jeroen J. A. Schellekens ◽  
Alexander C. A. P. Leenders ◽  
...  
Keyword(s):  
Early Diagnosis ◽  
Phase I ◽  
Real Time ◽  
Acute Phase ◽  
Phase Ii ◽  
Real Time Pcr ◽  
Q Fever ◽  
Serum Samples ◽  
Convalescent Phase ◽  
Acute Q Fever

ABSTRACT The world's largest Q fever outbreak is ongoing in The Netherlands with around 3,000 confirmed cases since the first half of 2007. Increased awareness has resulted in early referral of patients for diagnostics. An important drawback to serological diagnosis of acute Q fever is the lag phase in antibody response. Therefore, we evaluated the performance of a real-time PCR for detection of Coxiella burnetii DNA using serum samples from patients with acute Q fever. PCR, targeting IS1111, was retrospectively performed on acute-phase and follow-up convalescent-phase serum samples from 65 patients with acute Q fever as diagnosed by immunofluorescence assay. The results obtained by PCR were related to disease stage as defined by subsequent appearance of phase II IgM, phase II IgG, phase I IgM, and phase I IgG (IgM-II, IgG-II, IgM-I, and IgG-I, respectively) antibodies and time since onset of disease. In addition, we analyzed seronegative acute-phase serum samples from patients with inconclusive Q fever serology, because no convalescent-phase serum samples were available. PCR was scored positive in 49/50 (98%) seronegative sera, 9/10 (90%) sera with isolated IgM-II antibodies, 3/13 (23%) sera with IgM-II/IgG-II antibodies, 2/41 (5%) sera with IgM-II/IgG-II/IgM-I antibodies, 0/15 (0%) sera with IgM-II/IgG-II/IgM-I/IgG-I antibodies, and 0/1 (0%) serum sample with IgM-II/IgG-II/IgG-I antibodies. The latest time point after onset of disease in which C. burnetii DNA could be detected was at day 17. In patients with inconclusive Q fever serology, PCR was positive in 5/50 (10%) cases. We conclude that real-time PCR with serum samples is indispensable for early diagnosis of acute Q fever. C. burnetii DNA becomes undetectable in serum as the serological response develops.

Minimizing the risk of occupational Q fever exposure: A protocol for ensuring Coxiella burnetii–negative pregnant ewes are used for medical research

2020 ◽  
pp. 002367722096562
Author(s):  
Laura A Galganski ◽  
Benjamin A Keller ◽  
Connor Long ◽  
Kaeli J Yamashiro ◽  
Mennatalla S Hegazi ◽  
...  
Keyword(s):  
Amniotic Fluid ◽  
Phase I ◽  
Phase Ii ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Buffy Coat ◽  
Ovis Aries ◽  
Enzyme Linked Immunosorbent Assay ◽  
Research Facility ◽  
Increased Risk

Q fever is a worldwide zoonosis caused by Coxiella burnetii that can lead to abortion, endocarditis, and death in humans. Researchers utilizing parturient domestic ruminants, including sheep, have an increased risk of occupational exposure. This study evaluated the effectiveness of our screening protocol in eliminating C. burnetii–positive sheep from our facility. From August 2010 to May 2018, all ewes ( N  =  306) and select lambs ( N  =  272; ovis aries) were screened twice for C. burnetii utilizing a serum Phase I and Phase II antibody immunofluorescence assay (IFA). The first screen was performed by the vendor prior to breeding, and the second screen was performed on arrival to the research facility. Ewes that were positive on arrival screening were quarantined and retested using repeat IFA serology, enzyme-linked immunosorbent assay, buffy coat polymerase chain reaction (PCR), and amniotic fluid PCR. The overall individual seroprevalence of C. burnetii in the flocks tested by the vendor was 14.2%. Ewes with negative Phase I and Phase II IFA results were selected for transport to the research facility. Upon arrival to the facility, two (0.7%) ewes had positive Phase I IFA results. Repeat testing demonstrated seropositivity in one of these two ewes, though amniotic fluid PCR was negative in both. The repeat seropositive ewe was euthanized prior to use in a research protocol. No Q fever was reported among husbandry, laboratory or veterinary staff during the study period. Serologic testing for C. burnetii with IFA prior to transport and following arrival to a research facility limits potential exposure to research staff.

scholarly journals Evaluation of Commonly Used Serological Tests for Detection of Coxiella burnetii Antibodies in Well-Defined Acute and Follow-Up Sera

2012 ◽  
Vol 19 (7) ◽  
pp. 1110-1115 ◽  
Author(s):  
M. C. A. Wegdam-Blans ◽  
C. C. H. Wielders ◽  
J. Meekelenkamp ◽  
J. M. Korbeeck ◽  
T. Herremans ◽  
...  
Keyword(s):  
Phase Ii ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibodies ◽  
Serological Tests ◽  
Content Type ◽  
Acute Q Fever

ABSTRACTIn this study, we comparedCoxiella burnetiiIgG phase I, IgG phase II, and IgM phase II detection among a commercially available enzyme-linked immunosorbent assay (ELISA) (Virion/Serion), an indirect fluorescent antibody test (IFAT) (Focus Diagnostics), and a complement fixation test (CFT) (Virion/Serion). For this, we used a unique collection of acute- and convalescent-phase sera from 126 patients with acute Q fever diagnosed by positiveCoxiella burnetiiPCR of blood. We were able to establish a reliable date of onset of disease, since DNA is detectable within 2 weeks after the start of symptoms. In acute samples, att= 0, IFAT demonstrated IgM phase II antibodies in significantly more sera than did ELISA (31.8% versus 19.7%), although the portion of solitary IgM phase II was equal for IFAT and for ELISA (18.2% and 16.7%, respectively). Twelve months after the diagnosis of acute Q fever, 83.5% and 62.2% of the sera were still positive for IgM phase II with IFAT and ELISA, respectively. At 12 months IFAT IgG phase II showed the slowest decline. Therefore, definitive serological evidence of acute Q fever cannot be based on a single serum sample in areas of epidemicity and should involve measurement of both IgM and IgG antibodies in paired serum. Based on IgG phase II antibody detection in paired samples (at 0 and 3 months) from 62 patients, IFAT confirmed more cases than ELISA and CFT, but the differences were not statically significant (100% for IFAT, 95.2% for ELISA, and 96.8% for CFT). This study demonstrated that the three serological tests are equally effective in diagnosing acute Q fever within 3 months of start of symptoms. In follow-up sera, more IgG antibodies were detected by IFAT than by ELISA or CFT, making IFAT more suitable for prevaccination screening programs.

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