Detection of scrub typhus by real-time polymerase chain reaction and immunoglobulin M ELISA among patients with acute febrile illness
Background: Scrub typhus caused by Orientia tsutsugamushi is a vector-borne zoonotic infection endemic in several parts of the globe. The infection generally presents with fever and nonspecific clinical features but may lead to severe complications with a high mortality rate if untreated. Early diag...
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| creator | Saravanan, Nithiyanandan Rajendiran, Prashanth Sankar, Sathish Ramamurthy, Mageshbabu Sasimohan, Archana Vineeta, Vishnu Varghese, George Idikula, Mercy Jesudason, Mary Mangalakumar, Raja Nair, Aravindan Babujanarthanam, Ranganathan Nandagopal, Balaji Sridharan, Gopalan |
| description | Background: Scrub typhus caused by Orientia tsutsugamushi is a vector-borne zoonotic infection endemic in several parts of the globe. The infection generally presents with fever and nonspecific clinical features but may lead to severe complications with a high mortality rate if untreated. Early diagnosis and timely management are therefore important. Serological diagnosis such as Weil-Felix test, indirect immunofluorescence assay, immunoglobulin (Ig) M/IgG ELISA, and rapid antibody detection assays are either less sensitive or laborious. Molecular detection by polymerase chain reaction (PCR) targeting specific gene targets of O. tsutsugamushi is warranted. Materials and Methods: We developed a real-time PCR assay targeting 47-KDa htrA gene for the specific diagnosis of the pathogen. The assay was evaluated in a buffy coat from whole blood or serum samples collected from patients presenting with acute febrile illness. Randomly selected samples were also tested for IgM by commercial IgM ELISA assay. Results: The real-time PCR assay was able to detect |
| doi_str_mv | 10.4103/jnsbm.JNSBM_156_19 |
| format | Article |
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The infection generally presents with fever and nonspecific clinical features but may lead to severe complications with a high mortality rate if untreated. Early diagnosis and timely management are therefore important. Serological diagnosis such as Weil-Felix test, indirect immunofluorescence assay, immunoglobulin (Ig) M/IgG ELISA, and rapid antibody detection assays are either less sensitive or laborious. Molecular detection by polymerase chain reaction (PCR) targeting specific gene targets of O. tsutsugamushi is warranted. Materials and Methods: We developed a real-time PCR assay targeting 47-KDa htrA gene for the specific diagnosis of the pathogen. The assay was evaluated in a buffy coat from whole blood or serum samples collected from patients presenting with acute febrile illness. Randomly selected samples were also tested for IgM by commercial IgM ELISA assay. Results: The real-time PCR assay was able to detect <1 genome copy per the PCR input and specific to O. tsutsugamushi on heterologous pathogens testing. The samples were negative by real-time PCR and 13 samples were positive by IgM ELISA. This study found a relatively low prevalence of scrub typhus in the population. Conclusion: The assay developed in this study could be a useful diagnostic tool for the detection of O. tsutsugamushi in clinical samples. The study also indicated the need for a wide epidemiological survey that could help determine appropriate health measures including treatment and prevention.</description><identifier>ISSN: 0976-9668</identifier><identifier>EISSN: 2229-7707</identifier><identifier>DOI: 10.4103/jnsbm.JNSBM_156_19</identifier><language>eng</language><publisher>Mumbai: Wolters Kluwer India Pvt. Ltd</publisher><subject>Antibodies ; Buffy coat ; Deoxyribonucleic acid ; Diagnosis ; Diseases ; DNA ; EDTA ; Enzyme-linked immunosorbent assay ; Epidemiology ; Fever ; Fluorescent antibody technique ; Gene amplification ; Genomes ; Health aspects ; Health care ; Hospitals ; HtrA gene ; Illnesses ; Immunofluorescence ; Immunoglobulin G ; Immunoglobulin M ; Immunoglobulins ; Infection ; Infections ; Males ; Orientia tsutsugamushi ; Pathogens ; Patients ; Polymerase chain reaction ; Preventive medicine ; Public health ; Scrub typhus ; Time ; Typhus ; Zoonoses</subject><ispartof>Journal of natural science, biology and medicine, 2020-01, Vol.11 (1), p.66-71</ispartof><rights>COPYRIGHT 2020 Medknow Publications and Media Pvt. Ltd.</rights><rights>2020. This work is published under https://creativecommons.org/licenses/by-nc-sa/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c385o-aa128907f880915a8c0132791536c4b4c044add65a74c33939a7d73c87ecf8fb3</citedby><cites>FETCH-LOGICAL-c385o-aa128907f880915a8c0132791536c4b4c044add65a74c33939a7d73c87ecf8fb3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,4024,27923,27924,27925</link.rule.ids></links><search><creatorcontrib>Saravanan, Nithiyanandan</creatorcontrib><creatorcontrib>Rajendiran, Prashanth</creatorcontrib><creatorcontrib>Sankar, Sathish</creatorcontrib><creatorcontrib>Ramamurthy, Mageshbabu</creatorcontrib><creatorcontrib>Sasimohan, Archana</creatorcontrib><creatorcontrib>Vineeta, Vishnu</creatorcontrib><creatorcontrib>Varghese, George</creatorcontrib><creatorcontrib>Idikula, Mercy</creatorcontrib><creatorcontrib>Jesudason, Mary</creatorcontrib><creatorcontrib>Mangalakumar, Raja</creatorcontrib><creatorcontrib>Nair, Aravindan</creatorcontrib><creatorcontrib>Babujanarthanam, Ranganathan</creatorcontrib><creatorcontrib>Nandagopal, Balaji</creatorcontrib><creatorcontrib>Sridharan, Gopalan</creatorcontrib><title>Detection of scrub typhus by real-time polymerase chain reaction and immunoglobulin M ELISA among patients with acute febrile illness</title><title>Journal of natural science, biology and medicine</title><description>Background: Scrub typhus caused by Orientia tsutsugamushi is a vector-borne zoonotic infection endemic in several parts of the globe. The infection generally presents with fever and nonspecific clinical features but may lead to severe complications with a high mortality rate if untreated. Early diagnosis and timely management are therefore important. Serological diagnosis such as Weil-Felix test, indirect immunofluorescence assay, immunoglobulin (Ig) M/IgG ELISA, and rapid antibody detection assays are either less sensitive or laborious. Molecular detection by polymerase chain reaction (PCR) targeting specific gene targets of O. tsutsugamushi is warranted. Materials and Methods: We developed a real-time PCR assay targeting 47-KDa htrA gene for the specific diagnosis of the pathogen. The assay was evaluated in a buffy coat from whole blood or serum samples collected from patients presenting with acute febrile illness. Randomly selected samples were also tested for IgM by commercial IgM ELISA assay. Results: The real-time PCR assay was able to detect <1 genome copy per the PCR input and specific to O. tsutsugamushi on heterologous pathogens testing. The samples were negative by real-time PCR and 13 samples were positive by IgM ELISA. This study found a relatively low prevalence of scrub typhus in the population. Conclusion: The assay developed in this study could be a useful diagnostic tool for the detection of O. tsutsugamushi in clinical samples. The study also indicated the need for a wide epidemiological survey that could help determine appropriate health measures including treatment and prevention.</description><subject>Antibodies</subject><subject>Buffy coat</subject><subject>Deoxyribonucleic acid</subject><subject>Diagnosis</subject><subject>Diseases</subject><subject>DNA</subject><subject>EDTA</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Epidemiology</subject><subject>Fever</subject><subject>Fluorescent antibody technique</subject><subject>Gene amplification</subject><subject>Genomes</subject><subject>Health aspects</subject><subject>Health care</subject><subject>Hospitals</subject><subject>HtrA gene</subject><subject>Illnesses</subject><subject>Immunofluorescence</subject><subject>Immunoglobulin G</subject><subject>Immunoglobulin M</subject><subject>Immunoglobulins</subject><subject>Infection</subject><subject>Infections</subject><subject>Males</subject><subject>Orientia tsutsugamushi</subject><subject>Pathogens</subject><subject>Patients</subject><subject>Polymerase chain reaction</subject><subject>Preventive medicine</subject><subject>Public health</subject><subject>Scrub typhus</subject><subject>Time</subject><subject>Typhus</subject><subject>Zoonoses</subject><issn>0976-9668</issn><issn>2229-7707</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqNkt-KEzEUhwdRsOzuC3gV8MqLqclkJpm5rHV3rXQVrF6HTOZMm27-jEmG2gfwvXfWKloQMQkkcL4vhwO_LHtB8LwkmL7eu9ja-fsPmzd3glRMkOZJNiuKosk5x_xpNsMNZ3nDWP08u4pxj6dVTaegs-z7W0igkvYO-R5FFcYWpeOwGyNqjyiANHnSFtDgzdFCkBGQ2kntHksnTboOaWtH57fGt6OZanfoer3aLJC03m3RIJMGlyI66LRDUo0JUA9t0AaQNsZBjJfZs16aCFc_74vsy8315-W7fP3xdrVcrHNF68rnUpKibjDv6xo3pJK1woQWfHpSpsq2VLgsZdexSvJSUdrQRvKOU1VzUH3dt_Qie3n6dwj-6wgxib0fg5taioJyzgrGS_yb2koDQrvepyCV1VGJBSOcEEIJm6j5X6hpd2C18g76acBz4dWZMDEJvqWtHGMUq82n_2br2_U5W5xYFXyMAXoxBG1lOAqCxWNAxI-AiD8DMknLk3TwJkGI92Y8QBAWunvnD_8wBWPiV2boA1LSxbE</recordid><startdate>20200101</startdate><enddate>20200101</enddate><creator>Saravanan, Nithiyanandan</creator><creator>Rajendiran, Prashanth</creator><creator>Sankar, Sathish</creator><creator>Ramamurthy, Mageshbabu</creator><creator>Sasimohan, Archana</creator><creator>Vineeta, Vishnu</creator><creator>Varghese, George</creator><creator>Idikula, Mercy</creator><creator>Jesudason, Mary</creator><creator>Mangalakumar, Raja</creator><creator>Nair, Aravindan</creator><creator>Babujanarthanam, Ranganathan</creator><creator>Nandagopal, Balaji</creator><creator>Sridharan, Gopalan</creator><general>Wolters Kluwer India Pvt. 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G</topic><topic>Immunoglobulin M</topic><topic>Immunoglobulins</topic><topic>Infection</topic><topic>Infections</topic><topic>Males</topic><topic>Orientia tsutsugamushi</topic><topic>Pathogens</topic><topic>Patients</topic><topic>Polymerase chain reaction</topic><topic>Preventive medicine</topic><topic>Public health</topic><topic>Scrub typhus</topic><topic>Time</topic><topic>Typhus</topic><topic>Zoonoses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Saravanan, Nithiyanandan</creatorcontrib><creatorcontrib>Rajendiran, Prashanth</creatorcontrib><creatorcontrib>Sankar, Sathish</creatorcontrib><creatorcontrib>Ramamurthy, Mageshbabu</creatorcontrib><creatorcontrib>Sasimohan, Archana</creatorcontrib><creatorcontrib>Vineeta, Vishnu</creatorcontrib><creatorcontrib>Varghese, George</creatorcontrib><creatorcontrib>Idikula, Mercy</creatorcontrib><creatorcontrib>Jesudason, Mary</creatorcontrib><creatorcontrib>Mangalakumar, 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Edition</collection><collection>ProQuest Central China</collection><jtitle>Journal of natural science, biology and medicine</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Saravanan, Nithiyanandan</au><au>Rajendiran, Prashanth</au><au>Sankar, Sathish</au><au>Ramamurthy, Mageshbabu</au><au>Sasimohan, Archana</au><au>Vineeta, Vishnu</au><au>Varghese, George</au><au>Idikula, Mercy</au><au>Jesudason, Mary</au><au>Mangalakumar, Raja</au><au>Nair, Aravindan</au><au>Babujanarthanam, Ranganathan</au><au>Nandagopal, Balaji</au><au>Sridharan, Gopalan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of scrub typhus by real-time polymerase chain reaction and immunoglobulin M ELISA among patients with acute febrile illness</atitle><jtitle>Journal of natural science, biology and medicine</jtitle><date>2020-01-01</date><risdate>2020</risdate><volume>11</volume><issue>1</issue><spage>66</spage><epage>71</epage><pages>66-71</pages><issn>0976-9668</issn><eissn>2229-7707</eissn><abstract>Background: Scrub typhus caused by Orientia tsutsugamushi is a vector-borne zoonotic infection endemic in several parts of the globe. The infection generally presents with fever and nonspecific clinical features but may lead to severe complications with a high mortality rate if untreated. Early diagnosis and timely management are therefore important. Serological diagnosis such as Weil-Felix test, indirect immunofluorescence assay, immunoglobulin (Ig) M/IgG ELISA, and rapid antibody detection assays are either less sensitive or laborious. Molecular detection by polymerase chain reaction (PCR) targeting specific gene targets of O. tsutsugamushi is warranted. Materials and Methods: We developed a real-time PCR assay targeting 47-KDa htrA gene for the specific diagnosis of the pathogen. The assay was evaluated in a buffy coat from whole blood or serum samples collected from patients presenting with acute febrile illness. Randomly selected samples were also tested for IgM by commercial IgM ELISA assay. Results: The real-time PCR assay was able to detect <1 genome copy per the PCR input and specific to O. tsutsugamushi on heterologous pathogens testing. The samples were negative by real-time PCR and 13 samples were positive by IgM ELISA. This study found a relatively low prevalence of scrub typhus in the population. Conclusion: The assay developed in this study could be a useful diagnostic tool for the detection of O. tsutsugamushi in clinical samples. The study also indicated the need for a wide epidemiological survey that could help determine appropriate health measures including treatment and prevention.</abstract><cop>Mumbai</cop><pub>Wolters Kluwer India Pvt. Ltd</pub><doi>10.4103/jnsbm.JNSBM_156_19</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Journal of natural science, biology and medicine, 2020-01, Vol.11 (1), p.66-71 |
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| subjects | Antibodies Buffy coat Deoxyribonucleic acid Diagnosis Diseases DNA EDTA Enzyme-linked immunosorbent assay Epidemiology Fever Fluorescent antibody technique Gene amplification Genomes Health aspects Health care Hospitals HtrA gene Illnesses Immunofluorescence Immunoglobulin G Immunoglobulin M Immunoglobulins Infection Infections Males Orientia tsutsugamushi Pathogens Patients Polymerase chain reaction Preventive medicine Public health Scrub typhus Time Typhus Zoonoses |
| title | Detection of scrub typhus by real-time polymerase chain reaction and immunoglobulin M ELISA among patients with acute febrile illness |
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