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PLoS neglected tropical diseases2013; 7(5); e2231; doi: 10.1371/journal.pntd.0002231

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.
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Publication Date: 2013-05-16 PubMed ID: 23696915PubMed Central: PMC3656153DOI: 10.1371/journal.pntd.0002231Google Scholar: Lookup
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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

PLoS neglected tropical diseases
ISSN: 1935-2735
NlmUniqueID: 101291488
Country: United States
Language: English
Volume: 7
Issue: 5
Pages: e2231
PII: e2231

Researcher Affiliations

Pan, Lei
  • Dept. of Rickettsiology, National Institute of Communicable Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
Zhang, Lijuan
    Fan, Desheng
      Zhang, Xiuchun
        Liu, Hong
          Lu, Qunying
            Xu, Qiyi

              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.

              References

              This article includes 17 references
              1. Angelakis E, Raoult D. Q Fever.. Vet Microbiol 2010 Jan 27;140(3-4):297-309.
                pubmed: 19875249doi: 10.1016/j.vetmic.2009.07.016google scholar: lookup
              2. Raoult D. Chronic Q fever: expert opinion versus literature analysis and consensus.. J Infect 2012 Aug;65(2):102-8.
                pubmed: 22537659doi: 10.1016/j.jinf.2012.04.006google scholar: lookup
              3. Raoult D. Reemergence of Q fever after 11 September 2001.. Clin Infect Dis 2009 Mar 1;48(5):558-9.
                pubmed: 19191639doi: 10.1086/596706google scholar: lookup
              4. Graves S, Stenos J. Rickettsioses in Australia.. Ann N Y Acad Sci 2009 May;1166:151-5.
                pubmed: 19538275doi: 10.1111/j.1749-6632.2009.04530.xgoogle scholar: lookup
              5. Walker DH(1996) Rickettsiae.In Baron S, editor. Medical Microbiology. 4th edition. Galveston (TX): University of Texas Medical Branch at Galveston;
              6. Wen BH, Yu SR, Yu GQ, Li QJ, Zhang X. Analysis of proteins and lipopolysaccharides from Chinese isolates of Coxiella burnetii with monoclonal antibodies.. Acta Virol 1991 Nov;35(6):538-44.
                pubmed: 1687637
              7. Zhang LJ, Fu XP, Fan MY. Epidemiology and research situation of rickettsia and reckettsiosis in China. J Trop Dis Parasitol 3: 37–41.
              8. Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T. Loop-mediated isothermal amplification of DNA.. Nucleic Acids Res 2000 Jun 15;28(12):E63.
                pmc: PMC102748pubmed: 10871386doi: 10.1093/nar/28.12.e63google scholar: lookup
              9. Fan DS, Xu QY, Yu HL, Peng Y, Wang SW. Epidemiological surveys of Q Fever in YiLi regions of Xinjiang Province. Dis Survil 26 1: 15–17.
              10. Ya HX, Zhang LJ. Quantitative detection of Coxiella burneti by real time PCR. China Trop Med 11 8: 916–917.
              11. Roux V, Raoult D. Phylogenetic analysis of the genus Rickettsia by 16S rDNA sequencing.. Res Microbiol 1995 Jun;146(5):385-96.
                pubmed: 8525055doi: 10.1016/0923-2508(96)80284-1google scholar: lookup
              12. Fenollar F, Fournier PE, Raoult D. Molecular detection of Coxiella burnetii in the sera of patients with Q fever endocarditis or vascular infection.. J Clin Microbiol 2004 Nov;42(11):4919-24.
                pmc: PMC525147pubmed: 15528674doi: 10.1128/jcm.42.11.4919-4924.2004google scholar: lookup
              13. Hou MY, Hung MN, Lin PS, Wang YC, Lin CC, Shu PY, Shih WY, Wu HS, Lin LJ. Use of a single-tube nested real-time PCR assay to facilitate the early diagnosis of acute Q fever.. Jpn J Infect Dis 2011;64(2):161-2.
                pubmed: 21519134
              14. Ogawa M, Setiyono A, Sato K, Cai Y, Shiga S, Kishimoto T. Evaluation of PCR and nested PCR assays currently used for detection of Coxiella burnetii in Japan.. Southeast Asian J Trop Med Public Health 2004 Dec;35(4):852-5.
                pubmed: 15916080
              15. Schneeberger PM, Hermans MH, van Hannen EJ, Schellekens JJ, Leenders AC, Wever PC. Real-time PCR with serum samples is indispensable for early diagnosis of acute Q fever.. Clin Vaccine Immunol 2010 Feb;17(2):286-90.
                pmc: PMC2815520pubmed: 20032219doi: 10.1128/cvi.00454-09google scholar: lookup
              16. Klee SR, Tyczka J, Ellerbrok H, Franz T, Linke S, Baljer G, Appel B. Highly sensitive real-time PCR for specific detection and quantification of Coxiella burnetii.. BMC Microbiol 2006 Jan 19;6:2.
                pmc: PMC1360083pubmed: 16423303doi: 10.1186/1471-2180-6-2google scholar: lookup
              17. Xu ZY. Epidemiology of Parasites and Vectors. 253–258.

              Citations

              This article has been cited 14 times.
              1. Miller HK, Priestley RA, Kersh GJ. Q Fever: A troubling disease and a challenging diagnosis. Clin Microbiol Newsl 2021 Aug;43(13):109-118.
                doi: 10.1016/j.clinmicnews.2021.06.003pubmed: 37701818google scholar: lookup
              2. Ullah Q, Jamil T, Saqib M, Iqbal M, Neubauer H. Q Fever-A Neglected Zoonosis. Microorganisms 2022 Jul 28;10(8).
                doi: 10.3390/microorganisms10081530pubmed: 36013948google scholar: lookup
              3. Ni J, Lin H, Xu X, Ren Q, Aizezi M, Luo J, Luo Y, Ma Z, Chen Z, Tan Y, Guo J, Liu W, Qu Z, Wu Z, Wang J, Li Y, Guan G, Luo J, Yin H, Liu G. Coxiella burnetii is widespread in ticks (Ixodidae) in the Xinjiang areas of China. BMC Vet Res 2020 Aug 28;16(1):317.
                doi: 10.1186/s12917-020-02538-6pubmed: 32859190google scholar: lookup
              4. Khademi P, Ownagh A, Ataei B, Kazemnia A, Eydi J, Khalili M, M M, Mardani K. Molecular detection of Coxiella burnetii in horse sera in Iran. Comp Immunol Microbiol Infect Dis 2020 Oct;72:101521.
                doi: 10.1016/j.cimid.2020.101521pubmed: 32721772google scholar: lookup
              5. Sheikh N, Kumar S, Sharma HK, Bhagyawant SS, Thavaselvam D. Development of a Rapid and Sensitive Colorimetric Loop-Mediated Isothermal Amplification Assay: A Novel Technology for the Detection of Coxiella burnetii From Minimally Processed Clinical Samples. Front Cell Infect Microbiol 2020;10:127.
                doi: 10.3389/fcimb.2020.00127pubmed: 32432047google scholar: lookup
              6. Bae M, Jin CE, Park JH, Kim MJ, Chong YP, Lee SO, Choi SH, Kim YS, Woo JH, Shin Y, Kim SH. Diagnostic usefulness of molecular detection of Coxiella burnetii from blood of patients with suspected acute Q fever. Medicine (Baltimore) 2019 Jun;98(23):e15724.
                doi: 10.1097/MD.0000000000015724pubmed: 31169672google scholar: lookup
              7. Noden BH, Martin J, Carrillo Y, Talley JL, Ochoa-Corona FM. Development of a loop-mediated isothermal amplification (LAMP) assay for rapid screening of ticks and fleas for spotted fever group rickettsia. PLoS One 2018;13(2):e0192331.
                doi: 10.1371/journal.pone.0192331pubmed: 29390021google scholar: lookup
              8. Zhang Y, Yao Y, Du W, Wu K, Xu W, Lin M, Tan H, Li J. Development of loop-mediated isothermal amplification with Plasmodium falciparum unique genes for molecular diagnosis of human malaria. Pathog Glob Health 2017 Jul;111(5):247-255.
                doi: 10.1080/20477724.2017.1347379pubmed: 28683669google scholar: lookup
              9. Hu S, Niu L, Luo L, Song X, Sun J, Liu Q. Rapid, Sensitive Detection of Bartonella quintana by Loop-Mediated Isothermal Amplification of the groEL Gene. Int J Mol Sci 2016 Dec 1;17(12).
                doi: 10.3390/ijms17121902pubmed: 27916953google scholar: lookup
              10. Seo MG, Lee SH, VanBik D, Ouh IO, Yun SH, Choi E, Park YS, Lee SE, Kim JW, Cho GJ, Kwon OD, Kwak D. Detection and Genotyping of Coxiella burnetii and Coxiella-Like Bacteria in Horses in South Korea. PLoS One 2016;11(5):e0156710.
                doi: 10.1371/journal.pone.0156710pubmed: 27244230google scholar: lookup
              11. Dal Pozzo F, Renaville B, Martinelle L, Renaville R, Thys C, Smeets F, Kirschvink N, Grégoire F, Delooz L, Czaplicki G, Saegerman C. Single Nucleotide Polymorphism Genotyping and Distribution of Coxiella burnetii Strains from Field Samples in Belgium. Appl Environ Microbiol 2016 Jan 1;82(1):81-6.
                doi: 10.1128/AEM.02799-15pubmed: 26475104google scholar: lookup
              12. Angkasekwinai N, Atkins EH, Johnson RN, Grieco JP, Ching WM, Chao CC. Rapid and sensitive detection of Bartonella bacilliformis in experimentally infected sand flies by loop-mediated isothermal amplification (LAMP) of the Pap31 gene. PLoS Negl Trop Dis 2014 Dec;8(12):e3342.
                doi: 10.1371/journal.pntd.0003342pubmed: 25522230google scholar: lookup
              13. El-Mahallawy HS, Lu G, Kelly P, Xu D, Li Y, Fan W, Wang C. Q fever in China: a systematic review, 1989-2013. Epidemiol Infect 2015 Mar;143(4):673-81.
                doi: 10.1017/S0950268814002593pubmed: 25274488google scholar: lookup
              14. Dahmani A, Salhi O, Nabi M, Boucif A, Mekroud M, Touhami NAK, Ouchene N. Abortive diseases in horses and donkeys in Algeria: a systematic review. Trop Anim Health Prod 2026 Jan 5;58(1):30.
                doi: 10.1007/s11250-025-04843-4pubmed: 41489843google scholar: lookup
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