Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene.
BACKGROUND: 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 disea...
Published in: | PLoS Neglected Tropical Diseases |
---|---|
Main Authors: | , , , , , , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Public Library of Science (PLoS)
2013
|
Subjects: | |
Online Access: | https://doi.org/10.1371/journal.pntd.0002231 https://doaj.org/article/10064e90630f4dcb91f6925360f8bc59 |
id |
ftdoajarticles:oai:doaj.org/article:10064e90630f4dcb91f6925360f8bc59 |
---|---|
record_format |
openpolar |
spelling |
ftdoajarticles:oai:doaj.org/article:10064e90630f4dcb91f6925360f8bc59 2023-05-15T15:18:26+02:00 Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene. Lei Pan Lijuan Zhang Desheng Fan Xiuchun Zhang Hong Liu Qunying Lu Qiyi Xu 2013-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0002231 https://doaj.org/article/10064e90630f4dcb91f6925360f8bc59 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC3656153?pdf=render https://doaj.org/toc/1935-2735 1935-2735 doi:10.1371/journal.pntd.0002231 https://doaj.org/article/10064e90630f4dcb91f6925360f8bc59 PLoS Neglected Tropical Diseases, Vol 7, Iss 5, p e2231 (2013) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2013 ftdoajarticles https://doi.org/10.1371/journal.pntd.0002231 2022-12-30T22:38:08Z BACKGROUND: 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 ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLoS Neglected Tropical Diseases 7 5 e2231 |
institution |
Open Polar |
collection |
Directory of Open Access Journals: DOAJ Articles |
op_collection_id |
ftdoajarticles |
language |
English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
spellingShingle |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Lei Pan Lijuan Zhang Desheng Fan Xiuchun Zhang Hong Liu Qunying Lu Qiyi Xu Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
BACKGROUND: 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 ... |
format |
Article in Journal/Newspaper |
author |
Lei Pan Lijuan Zhang Desheng Fan Xiuchun Zhang Hong Liu Qunying Lu Qiyi Xu |
author_facet |
Lei Pan Lijuan Zhang Desheng Fan Xiuchun Zhang Hong Liu Qunying Lu Qiyi Xu |
author_sort |
Lei Pan |
title |
Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene. |
title_short |
Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene. |
title_full |
Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene. |
title_fullStr |
Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene. |
title_full_unstemmed |
Rapid, simple and sensitive detection of Q fever by loop-mediated isothermal amplification of the htpAB gene. |
title_sort |
rapid, simple and sensitive detection of q fever by loop-mediated isothermal amplification of the htpab gene. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2013 |
url |
https://doi.org/10.1371/journal.pntd.0002231 https://doaj.org/article/10064e90630f4dcb91f6925360f8bc59 |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 7, Iss 5, p e2231 (2013) |
op_relation |
http://europepmc.org/articles/PMC3656153?pdf=render https://doaj.org/toc/1935-2735 1935-2735 doi:10.1371/journal.pntd.0002231 https://doaj.org/article/10064e90630f4dcb91f6925360f8bc59 |
op_doi |
https://doi.org/10.1371/journal.pntd.0002231 |
container_title |
PLoS Neglected Tropical Diseases |
container_volume |
7 |
container_issue |
5 |
container_start_page |
e2231 |
_version_ |
1766348637578723328 |