TY - JOUR
T1 - Development of three quantitative real-time PCR assays for the detection of Rickettsia raoultii, Rickettsia slovaca, and Rickettsia aeschlimannii and their validation with ticks from the country of Georgia and the Republic of Azerbaijan
AU - Jiang, Ju
AU - You, Brian J.
AU - Liu, Evan
AU - Apte, Anisha
AU - Yarina, Tamasin R.
AU - Myers, Todd E.
AU - Lee, John S.
AU - Francesconi, Stephen C.
AU - O'Guinn, Monica L.
AU - Tsertsvadze, Nikoloz
AU - Vephkhvadze, Nino
AU - Babuadze, Giorgi
AU - Sidamonidze, Ketevan
AU - Kokhreidze, Maka
AU - Donduashvili, Marina
AU - Onashvili, Tinatin
AU - Ismayilov, Afrail
AU - Agayev, Nigar
AU - Aliyev, Mubariz
AU - Muttalibov, Nizam
AU - Richards, Allen L.
N1 - Funding Information:
Funding provided by the Defense Threat Reduction Agency , Work unit # B0017 60000.000.0.B0017. The views expressed in this article are those of the author and do not necessarily reflect the official policy or position of the Dept. of the Navy, Dept. of Defense, nor the U.S. Government. Authors, as employees of the U.S. Government, conducted the work as part of their official duties. Title 17 U.S.C. §105 provides that ‘Copyright protection under this title is not available for any work of the United States Government.’ Title 17 U.S.C. §101 defines a U.S. Government work as a work prepared by an employee of the U.S. Government as part of the person's official duties.
PY - 2012/12
Y1 - 2012/12
N2 - A previous surveillance study of human pathogens within ticks collected in the country of Georgia showed a relatively high infection rate for Rickettsia raoultii, R. slovaca, and R. aeschlimannii. These 3 spotted fever group rickettsiae are human pathogens: R. raoultii and R. slovaca cause tick-borne lymphadenopathy (TIBOLA), and R. aeschlimannii causes an infection characterized by fever and maculopapular rash. Three quantitative real-time polymerase chain reaction (qPCR) assays, Rraoul, Rslov, and Raesch were developed and optimized to detect R. raoultii, R. slovaca, and R. aeschlimannii, respectively, by targeting fragments of the outer membrane protein B gene (ompB) using species-specific molecular beacon or TaqMan probes. The 3 qPCR assays showed 100% specificity when tested against a rickettsiae DNA panel (n= 20) and a bacteria DNA panel (n= 12). The limit of detection was found to be at least 3 copies per reaction for all assays. Validation of the assays using previously investigated tick nucleic acid preparations, which included Rickettsia-free tick samples, tick samples that contain R. raoultii, R. slovaca, R. aeschlimannii, and other Rickettsia spp., gave 100% sensitivity for all 3 qPCR assays. In addition, a total of 65 tick nucleic acid preparations (representing 259 individual ticks) collected from the country of Georgia and the Republic of Azerbaijan in 2009 was tested using the 3 qPCR assays. R. raoultii, R. slovaca, and R. aeschlimannii were not detected in any ticks (n= 31) from the Republic of Azerbaijan, but in the ticks from the country of Georgia (n= 228) the minimal infection rate for R. raoultii and R. slovaca in Dermacentor marginatus was 10% and 4%, respectively, and for R. aeschlimannii in Haemaphysalis sulcata and Hyalomma spp. it was 1.9% and 20%, respectively.
AB - A previous surveillance study of human pathogens within ticks collected in the country of Georgia showed a relatively high infection rate for Rickettsia raoultii, R. slovaca, and R. aeschlimannii. These 3 spotted fever group rickettsiae are human pathogens: R. raoultii and R. slovaca cause tick-borne lymphadenopathy (TIBOLA), and R. aeschlimannii causes an infection characterized by fever and maculopapular rash. Three quantitative real-time polymerase chain reaction (qPCR) assays, Rraoul, Rslov, and Raesch were developed and optimized to detect R. raoultii, R. slovaca, and R. aeschlimannii, respectively, by targeting fragments of the outer membrane protein B gene (ompB) using species-specific molecular beacon or TaqMan probes. The 3 qPCR assays showed 100% specificity when tested against a rickettsiae DNA panel (n= 20) and a bacteria DNA panel (n= 12). The limit of detection was found to be at least 3 copies per reaction for all assays. Validation of the assays using previously investigated tick nucleic acid preparations, which included Rickettsia-free tick samples, tick samples that contain R. raoultii, R. slovaca, R. aeschlimannii, and other Rickettsia spp., gave 100% sensitivity for all 3 qPCR assays. In addition, a total of 65 tick nucleic acid preparations (representing 259 individual ticks) collected from the country of Georgia and the Republic of Azerbaijan in 2009 was tested using the 3 qPCR assays. R. raoultii, R. slovaca, and R. aeschlimannii were not detected in any ticks (n= 31) from the Republic of Azerbaijan, but in the ticks from the country of Georgia (n= 228) the minimal infection rate for R. raoultii and R. slovaca in Dermacentor marginatus was 10% and 4%, respectively, and for R. aeschlimannii in Haemaphysalis sulcata and Hyalomma spp. it was 1.9% and 20%, respectively.
KW - Quantitative real-time PCR assays
KW - R. aeschlimannii
KW - R. raoultii
KW - R. slovaca
KW - The country of Georgia
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U2 - 10.1016/j.ttbdis.2012.10.004
DO - 10.1016/j.ttbdis.2012.10.004
M3 - Article
C2 - 23182543
AN - SCOPUS:84870421990
SN - 1877-959X
VL - 3
SP - 327
EP - 331
JO - Ticks and Tick-borne Diseases
JF - Ticks and Tick-borne Diseases
IS - 5-6
ER -