Detection and distribution of Sca autotransporter protein antigens in diverse isolates of Orientia tsutsugamushi.
Orientia tsutsugamushi (Ots) frequently causes severe scrub typhus infections in the Asia-Pacific region. Korean investigators have demonstrated that Ots encodes five different autotransporter domain (ATD) proteins (ScaA-ScaE). ScaA functions as an adhesin and confers protective immunity in a lethal...
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ftdoajarticles:oai:doaj.org/article:40cf87c1ef804390b9e00e92fa96fec3 2023-05-15T15:18:28+02:00 Detection and distribution of Sca autotransporter protein antigens in diverse isolates of Orientia tsutsugamushi. Munegowda C Koralur Arunachalam Ramaiah Gregory A Dasch 2018-09-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0006784 https://doaj.org/article/40cf87c1ef804390b9e00e92fa96fec3 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC6168176?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0006784 https://doaj.org/article/40cf87c1ef804390b9e00e92fa96fec3 PLoS Neglected Tropical Diseases, Vol 12, Iss 9, p e0006784 (2018) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2018 ftdoajarticles https://doi.org/10.1371/journal.pntd.0006784 2022-12-31T11:18:38Z Orientia tsutsugamushi (Ots) frequently causes severe scrub typhus infections in the Asia-Pacific region. Korean investigators have demonstrated that Ots encodes five different autotransporter domain (ATD) proteins (ScaA-ScaE). ScaA functions as an adhesin and confers protective immunity in a lethal mouse model of Ots infection. Specific antibodies are detected against ScaA and ScaC in Korean scrub typhus patients. However, there is limited data on the distribution of the Sca protein genes in diverse isolates of Ots. By BLAST analysis with the conserved beta barrel autotransporter domain (ATD) regions of the sca proteins, we discovered a sixth gene scaF among 3 of 10 new partial Ots genome sequences available at NCBI GenBank (Sido, Karp, AFSC7). We designed two to seven specific TaqMan assays to detect the ATD for each of the six sca genes. The TaqMan assays among those for each sca gene which gave the greatest sensitivity and linearity with DNA log dilutions were then used for screening DNAs from Ots isolates grown in L929 mouse cells for sca genes. The sca prevalence survey was performed for all six sca genes with 178 DNAs from isolates from 12 countries. The survey results were confirmed by conventional PCR with primers from conserved regions of the passenger domains (PD) and ATD of the sca proteins. The ATD was highly conserved between the DNAs of different genotypes compared to the sca PD but each TaqMan assay was sca specific. The percentage positivity for 56 kDa and scaA genes in the 178 DNAs using Ha primers was 59.6% and 62.4%, respectively. Our scaA conventional ATD PCR assay was positive in 98.3% but scaA was present in all 178 DNAs (100%) by ATD TaqMan. scaB, scaC, scaD, scaE and scaF were detected in 33.7%, 97.8%, 93.8%, 97.2% and 43.3% isolates by ATD TaqMan, respectively. The ATDs of Ots sca genes are thus sufficiently conserved between different genotypes for molecular assay design. Four sca genes are widely distributed among diverse Ots isolates from diverse geographical areas. scaB and scaF ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic Pacific PLOS Neglected Tropical Diseases 12 9 e0006784 |
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English |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Munegowda C Koralur Arunachalam Ramaiah Gregory A Dasch Detection and distribution of Sca autotransporter protein antigens in diverse isolates of Orientia tsutsugamushi. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Orientia tsutsugamushi (Ots) frequently causes severe scrub typhus infections in the Asia-Pacific region. Korean investigators have demonstrated that Ots encodes five different autotransporter domain (ATD) proteins (ScaA-ScaE). ScaA functions as an adhesin and confers protective immunity in a lethal mouse model of Ots infection. Specific antibodies are detected against ScaA and ScaC in Korean scrub typhus patients. However, there is limited data on the distribution of the Sca protein genes in diverse isolates of Ots. By BLAST analysis with the conserved beta barrel autotransporter domain (ATD) regions of the sca proteins, we discovered a sixth gene scaF among 3 of 10 new partial Ots genome sequences available at NCBI GenBank (Sido, Karp, AFSC7). We designed two to seven specific TaqMan assays to detect the ATD for each of the six sca genes. The TaqMan assays among those for each sca gene which gave the greatest sensitivity and linearity with DNA log dilutions were then used for screening DNAs from Ots isolates grown in L929 mouse cells for sca genes. The sca prevalence survey was performed for all six sca genes with 178 DNAs from isolates from 12 countries. The survey results were confirmed by conventional PCR with primers from conserved regions of the passenger domains (PD) and ATD of the sca proteins. The ATD was highly conserved between the DNAs of different genotypes compared to the sca PD but each TaqMan assay was sca specific. The percentage positivity for 56 kDa and scaA genes in the 178 DNAs using Ha primers was 59.6% and 62.4%, respectively. Our scaA conventional ATD PCR assay was positive in 98.3% but scaA was present in all 178 DNAs (100%) by ATD TaqMan. scaB, scaC, scaD, scaE and scaF were detected in 33.7%, 97.8%, 93.8%, 97.2% and 43.3% isolates by ATD TaqMan, respectively. The ATDs of Ots sca genes are thus sufficiently conserved between different genotypes for molecular assay design. Four sca genes are widely distributed among diverse Ots isolates from diverse geographical areas. scaB and scaF ... |
format |
Article in Journal/Newspaper |
author |
Munegowda C Koralur Arunachalam Ramaiah Gregory A Dasch |
author_facet |
Munegowda C Koralur Arunachalam Ramaiah Gregory A Dasch |
author_sort |
Munegowda C Koralur |
title |
Detection and distribution of Sca autotransporter protein antigens in diverse isolates of Orientia tsutsugamushi. |
title_short |
Detection and distribution of Sca autotransporter protein antigens in diverse isolates of Orientia tsutsugamushi. |
title_full |
Detection and distribution of Sca autotransporter protein antigens in diverse isolates of Orientia tsutsugamushi. |
title_fullStr |
Detection and distribution of Sca autotransporter protein antigens in diverse isolates of Orientia tsutsugamushi. |
title_full_unstemmed |
Detection and distribution of Sca autotransporter protein antigens in diverse isolates of Orientia tsutsugamushi. |
title_sort |
detection and distribution of sca autotransporter protein antigens in diverse isolates of orientia tsutsugamushi. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2018 |
url |
https://doi.org/10.1371/journal.pntd.0006784 https://doaj.org/article/40cf87c1ef804390b9e00e92fa96fec3 |
geographic |
Arctic Pacific |
geographic_facet |
Arctic Pacific |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 12, Iss 9, p e0006784 (2018) |
op_relation |
http://europepmc.org/articles/PMC6168176?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0006784 https://doaj.org/article/40cf87c1ef804390b9e00e92fa96fec3 |
op_doi |
https://doi.org/10.1371/journal.pntd.0006784 |
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PLOS Neglected Tropical Diseases |
container_volume |
12 |
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9 |
container_start_page |
e0006784 |
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1766348660007763968 |