Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene.
Abstract: Q fever is the most widespread zoonosis, and domestic animals are the most common sources of transmission. It is not only difficult to distinguish from other febrile diseases because of the lack of specific clinical manifestations in humans, but it is also difficult to identify the disease in C. burnetii-carrying animals because of the lack of identifiable features. Conventional serodiagnosis requires sera from the acute and convalescent stages of infection, which are unavailable at early diagnosis. Nested PCR and real-time PCR require equipment. In this study, we developed a Loop-Mediated Isothermal Amplification (LAMP) assay to identify C. burnetii rapidly and sensitively. Methods: A universal LAMP primer set was designed to detect the repeated sequence IS1111a of the htpAB gene of C. burnetii using PrimerExplorer V4 software. The sensitivity of the LAMP assay was evaluated using known quantities of recombined reference plasmids containing the targeted genes. The specificity of the developed LAMP assay was determined using 26 members of order Rickettsiae and 18 other common pathogens. The utility of the LAMP assay was further compared with real time PCR by the examination 24 blood samples including 6 confirmed and 18 probable Q fever cases, which diagnosed by IFA serological assessment and real time PCR. In addition, 126 animal samples from 4 provinces including 97 goats, 7 cattle, 18 horses, 3 marmots and 1 deer were compared by these two methods. Results: The limits of detection of the LAMP assay for the htpAB gene were 1 copy per reaction. The specificity of the LAMP assay was 100%, and no cross-reaction was observed among the bacteria used in the study. The positive rate of unknown febrile patients was 33.3%(95%CI 30.2%-36.4%) for the LAMP assay and 8.3%(95%CI 7.4%-9.2%) for the real time PCR(P<0.05). Similarly, the total positive rate of animals was 7.9%(95%CI 7.1%-8.7%) for the LAMP assay and 0.8%(95%CI 0.7%-0.9%)for the real time PCR(P<0.01). Using the developed LAMP assay, Q fever in the Yi Li area, Xinjiang Province, was confirmed. Conclusions: The LAMP assay is a potential tool to support the diagnosis of Q fever in humans and domestic animals in the field, especially in the rural areas of China, because of its rapid and sensitive detection without the aid of sophisticated equipment or a complicated protocol.
Publication Date: 2013-05-16 PubMed ID: 23696915PubMed Central: PMC3656153DOI: 10.1371/journal.pntd.0002231Google Scholar: Lookup
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- Evaluation Study
- Journal Article
- Research Support
- Non-U.S. Gov't
- Diagnosis
- Diagnostic Technique
- Disease control
- Disease Diagnosis
- Disease Etiology
- Disease Management
- Disease Outbreaks
- Disease Surveillance
- Disease Transmission
- Disease Treatment
- Domestic Animals
- Epidemiology
- Equine Health
- Infectious Disease
- Public Health
- Veterinary Medicine
- Veterinary Procedure
- Veterinary Research
- Veterinary Science
- Zoonotic Diseases
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
This research presents a more efficient testing method, Loop-Mediated Isothermal Amplification (LAMP), for detecting Q fever in humans and animals, especially in rural China. LAMP has been shown to be quicker, easier, and more sensitive compared to traditional PCR methods.
Methodology
- The team designed a LAMP primer set to identify the htpAB gene sequence of C. burnetii (which causes Q fever).
- This method was tested for sensitivity with known quantities of recombined reference plasmids containing the targeted genes.
- The specificity of the LAMP assay was tested against 26 different types of bacteria and 18 other common pathogens.
- The researchers also tested the LAMP assay against real time PCR on 24 blood samples, including 6 confirmed and 18 probable Q fever cases, and on 126 animal samples from 4 provinces in China.
Results
- LAMP was able to detect the htpAB gene at a concentration of just 1 copy per reaction, demonstrating high sensitivity.
- The test was 100% specific, with no cross-reaction recorded among the different types of bacteria used in the study.
- The rate of positive results was notably higher with LAMP compared to real time PCR.
- Using LAMP, the team was able to confirm cases of Q fever in the Yi Li area of Xinjiang Province.
Conclusion
- LAMP represents a valuable tool for diagnosing Q fever in humans and domestic animals, particularly in less developed rural areas.
- This method offers rapid and sensitive detection without the need for complex equipment or procedures.
Cite This Article
APA
Pan L, Zhang L, Fan D, Zhang X, Liu H, Lu Q, Xu Q.
(2013).
Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene.
PLoS Negl Trop Dis, 7(5), e2231.
https://doi.org/10.1371/journal.pntd.0002231 Publication
Researcher Affiliations
- Dept. of Rickettsiology, National Institute of Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
MeSH Terms
- Adolescent
- Adult
- Aged
- Animals
- Animals, Domestic
- Animals, Wild
- Bacteriological Techniques / methods
- China
- Coxiella burnetii / genetics
- Coxiella burnetii / isolation & purification
- DNA Primers / genetics
- DNA Transposable Elements
- DNA, Bacterial / genetics
- Female
- Humans
- Male
- Middle Aged
- Molecular Diagnostic Techniques / methods
- Nucleic Acid Amplification Techniques / methods
- Q Fever / diagnosis
- Q Fever / veterinary
- Sensitivity and Specificity
- Young Adult
Conflict of Interest Statement
The authors have declared that no competing interests exist.
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Citations
This article has been cited 13 times.- Miller HK, Priestley RA, Kersh GJ. Q Fever: A troubling disease and a challenging diagnosis.. Clin Microbiol Newsl 2021 Aug;43(13):109-118.
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