Diagnosis of Acute Q Fever by Detection of Coxiella burnetii DNA using Real-Time PCR, Employing a Commercial Genesig Easy Kit

Pradeep, Jothimani; Spellman, Stephen R.; Ambroise, Stanley; Gunasekaran, Dhandapany

doi:10.7860/jcdr/2017/31005.10606

Cited by 12 publications

(12 citation statements)

References 27 publications

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“…We evaluated the performance of RT-qPCR for the detection of C. burnetii- specific DNA targeting the IS1111 gene using serum samples from patients with acute Q fever. The use of molecular detection based on the IS1111 gene has 100% specificity [ 69 ]. Not all seropositive samples were positive by RT-PCR.…”

Section: Discussionmentioning

confidence: 99%

High prevalence of Coxiella burnetii infection in humans and livestock in Assiut, Egypt: A serological and molecular survey

Abbass

Selim

Sobhy³

et al. 2020

Vet World

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Background and Aim: Q fever is considered a neglected zoonotic disease and is caused by Coxiella burnetii. Very little information is available on C. burnetii infections in cattle, sheep, and goat populations in Egypt. The aim of this study was to identify the seroprevalence of C. burnetii in humans and livestock and to test for the presence of C. burnetii DNA in sera from seropositive animals and humans. Materials and Methods: Blood samples were collected from 160 apparently healthy farm animals and 120 patients from three hospitals of the Assiut Governorate throughout 2017/2018. These populations were tested for antibodies against C. burnetii phase II antigen by immunofluorescence assay [IFA]) and enzyme-linked immunosorbent assay (ELISA). Seropositive samples were subjected to real-time quantitative polymerase chain reaction (RT-qPCR). Results: The results of the IFA revealed C. burnetii seroprevalence rates of 45.3%, 56.0%, 45.7%, and 53.3% in cattle, sheep, goats, and humans, respectively. In humans, the seroprevalence rates were 52.1%, 30.4%, 37.5%, 74.1%, and 62.5% in patients with fever of unknown origin, influenza, kidney dialysis, hepatitis C virus, and hepatitis B virus, respectively. Likewise, by ELISA, the seroprevalence in bovine was 50.7%; sheep, 60.0%; goats, 51.4%; and humans, 55.0% (54.3%, 30.4%, 37.5%, 77.8%, and 62.5% in patients with fever of unknown origin, influenza, kidney dialysis, hepatitis C virus, and hepatitis B virus, respectively). RT-qPCR targeting the repetitive element IS1111 confirmed the presence of C. burnetii DNA. Conclusion: These results proved that apparently healthy cattle, sheep, and goats may be very important reservoirs of C. burnetii infection. In light of these data, the effect of Q fever on the replication of hepatitis virus remains unclear. Although hepatitis is one of the main aspects of acute Q fever, the influence of hepatitis on Q fever remains to be investigated. Q fever is not a reportable disease in Egypt, and clinical cases may rarely be recognized by the health-care system. Additional information on the epidemiology of C. burnetii in Egypt is warranted, including other associated problems such as the distribution of infections, pathologic hallmarks, and molecular typing.

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Section: Discussionmentioning

confidence: 99%

High prevalence of Coxiella burnetii infection in humans and livestock in Assiut, Egypt: A serological and molecular survey

Abbass

Selim

Sobhy³

et al. 2020

Vet World

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“…Our in-house Q fever PCR detection method seems to have higher sensitivity (81%) and relatively high specificity when compared to those described in the previous studies, although wide confidence intervals prevent us from drawing a firm conclusion. [6–8] It is worth noting that the administration of doxycycline before PCR testing might have affected the results in 3 patients with false-negative results. In this study, the Q fever PCR was performed on samples collected a median of 16 days following symptom onset.…”

Section: Discussionmentioning

confidence: 99%

“…[5] Although there are some reports of molecular detection of C. burnetii DNA in buffy coat or serum from patients with acute Q fever, the sensitivity of PCR testing was reported to be approximately 33.3% to 66.7%. [6–8] We thus developed an in-house PCR test for Q fever and evaluated its diagnostic performance for Q fever detection using blood from patients with suspected acute Q fever.…”

Section: Introductionmentioning

confidence: 99%

Diagnostic usefulness of molecular detection of Coxiella burnetii from blood of patients with suspected acute Q fever

Bae¹,

Jin

Park³

et al. 2019

Medicine

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Diagnosis of Q fever is difficult due to the lack of distinct clinical features that distinguish it from other febrile diseases. Serologic testing is the gold standard method for diagnosing Q fever, but antibody formation may not be detectable for 2 to 3 weeks from symptom onset, limiting early diagnosis. We thus evaluated the diagnostic utility of polymerase chain reaction (PCR) to detect Coxellia burnetii DNA in serum from patients with suspected acute Q fever. All adult patients with suspected acute Q fever were prospectively enrolled at a tertiary-care hospital from January 2016 through July 2018. Acute Q fever was diagnosed using clinical and laboratory criteria: fever with at least one other symptoms (myalgia, headache, pneumonia, or hepatitis) and single phase II immunoglobulin G (IgG) antibody titers ≥1:200 or immunoglobulin M (IgM) antibody titer ≥1:50 (probable), or a fourfold increase or seroconversion in phase II IgG antibody titers as measured by indirect immunofluorescence assays between paired samples (confirmed). We performed PCR targeting the transposase gene insertion element IS1111a of C. burnetii . Of the 35 patients with suspected acute Q fever, 16 (46%) were diagnosed with acute Q fever including 8 probable and 8 confirmed cases; the remaining 19 (54%) were diagnosed with other febrile diseases. The proportion of males diagnosed with Q fever was higher than those diagnosed with other febrile diseases (88% vs 44%, P = .03), but there were no other significant differences in clinical characteristics between the 2 groups. The Q fever PCR sensitivity was 81% (95% confidence interval [CI], 54–96), specificity was 90% (95% CI, 67–99), positive predictive value was 87% (95% CI, 63–96), and negative predictive value was 85% (95% CI, 67–94). Q fever PCR testing using blood from patients with suspected acute Q fever seems to be a rapid and useful test for early diagnosis of Q fever.

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“…The prerequisite for such a diagnostic is a target sequence that is specific for C. burnetii to exclude false positive results with other organisms and also conserved in all isolates to prevent false negative reactions. The target sequences used so far include the plasmid genes (QpH1, QpRS) (Willems et al, 1993), the singular chromosomal genes like com1, htpB, (Seshadri et al, 2003), icd (Klee et al, 2006), icmX, icmW, icmV, icmT, dotB (Morgan et al, 2010, dotA (Omsland et al, 2009;Cockrell et al, 2017), gyrA gene (Pradeep et al, 2017) or the transposase gene of insertion element IS1111, which is present in the genome of C. burnetii in multiple copies. Although qPCR quantification of the C. burnetii cells is very sensitive due to the presence of a multicopy number of this insertion element in the genome, the number of copies of this element in different isolates seem to be highly variable, which makes the quantification difficult (Klee et al, 2006).…”

Section: Resultsmentioning

confidence: 99%

Counting of viable C. burnetii cells by quantitative reverse transcription PCR using a recombinant plasmid (pCB-dotA) as a standard

Zúñiga-Navarrete¹,

Flores-Ramirez²,

Quevedo-Diaz³

et al. 2018

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are usually infected by inhalation of aerosols, contaminated by infectious particles derived from domestic animals, particularly after contact with parturient females and their birth products (Babudieri, 1959; Marrie, 1990). Q fever is a zoonosis with a worldwide distribution. Many species of mammals, birds, and ticks are reservoirs of C. burnetii in nature. The infection in an animal is most often latent, with persistent shedding of bacteria into the environment. However, in the female reproductive tract, high-level shedding occurs at the time of parturition, with millions of bacteria being released in the placenta. Treatment with antimicrobial agents is recommended for both acute and chronic Q fever. For acute Q fever, a variety of antibiotics have been used, but the member of a tetracycline group, doxycycline, is considered the medicament of choice. Doxycycline has been shown to result in a mean time to defervescence of 2-3 days, whereas untreated patients resolve the fever after a mean of 12.5 days. This antibiotic has shown effectivity also for therapy of chronic form, but it was demonstrated that the combination of doxycycline with chloroquine

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Diagnosis of Acute Q Fever by Detection of Coxiella burnetii DNA using Real-Time PCR, Employing a Commercial Genesig Easy Kit

Cited by 12 publications

References 27 publications

High prevalence of Coxiella burnetii infection in humans and livestock in Assiut, Egypt: A serological and molecular survey

High prevalence of Coxiella burnetii infection in humans and livestock in Assiut, Egypt: A serological and molecular survey

Diagnostic usefulness of molecular detection of Coxiella burnetii from blood of patients with suspected acute Q fever

Counting of viable C. burnetii cells by quantitative reverse transcription PCR using a recombinant plasmid (pCB-dotA) as a standard

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