Molecular identification of diarrheal Aeromonas using immuno magnetic polymerase chain reaction (IM-PCR) technique: a comparative study with conventional culture method

BACKGROUND: Aeromonas are ubiquitous bacteria causing many clinical conditions including acute diarrhea. Diarrheagenic Aeromonas harbors aerolysin gene secreting virulent enterotoxin, aerolysin. OBJECTIVES: To develop a molecular and immunological based method for detection of Aeromonas. METHODS: Di...

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Autores principales: Subbaram, Kannan, Gatasheh, Mansour K, Al Azzam, Khaldun M, Kannan, Hemalatha
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Makerere Medical School 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6794535/
https://www.ncbi.nlm.nih.gov/pubmed/31656487
http://dx.doi.org/10.4314/ahs.v19i2.27
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author Subbaram, Kannan
Gatasheh, Mansour K
Al Azzam, Khaldun M
Kannan, Hemalatha
author_facet Subbaram, Kannan
Gatasheh, Mansour K
Al Azzam, Khaldun M
Kannan, Hemalatha
author_sort Subbaram, Kannan
collection PubMed
description BACKGROUND: Aeromonas are ubiquitous bacteria causing many clinical conditions including acute diarrhea. Diarrheagenic Aeromonas harbors aerolysin gene secreting virulent enterotoxin, aerolysin. OBJECTIVES: To develop a molecular and immunological based method for detection of Aeromonas. METHODS: Diarrheal Aeromonas strains were identified from stool samples using culture, enterotoxicity testing using mice model. During immune magnetic polymerase chain reaction IM-PCR protocol, aerolysin specific antibodies were bound with immuno magnetic binding. Sensitivity and specificity tests for IM-PCR were conducted. RESULTS: There was high detection of Aeromonas using IM-PCR (12.4 %) technique when compared to low isolation with culture (5.1%). Our study confirmed that some strains of enterotoxic Aeromonas strains were uncultivable. Enterotoxicity tests on culture isolates revealed many strains were negative. IM-PCR detected high, (62/500) rate of identification of Aeromonas with aerolysin toxin gene. Aeromonas species identified after IM-PCR were A. hydrophila (40.3% ), A. veronii (17.7 %), A. caviae (14.5 %), A. trota (11.2 %), A. jandei (9.6 %) and A. schuberti (6.4%). All A. trota strains were undetected by cultivation. CONCLUSION: High sensitivity and specificity of IM-PCR are due to preparation of aerolysin antibodies and immuno magnetic binding, prior to PCR. Since diseases due to Aeromonas are increasingly reported, IM-PCR is recommended for detection from clinical specimens.
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spelling pubmed-67945352019-10-25 Molecular identification of diarrheal Aeromonas using immuno magnetic polymerase chain reaction (IM-PCR) technique: a comparative study with conventional culture method Subbaram, Kannan Gatasheh, Mansour K Al Azzam, Khaldun M Kannan, Hemalatha Afr Health Sci Articles BACKGROUND: Aeromonas are ubiquitous bacteria causing many clinical conditions including acute diarrhea. Diarrheagenic Aeromonas harbors aerolysin gene secreting virulent enterotoxin, aerolysin. OBJECTIVES: To develop a molecular and immunological based method for detection of Aeromonas. METHODS: Diarrheal Aeromonas strains were identified from stool samples using culture, enterotoxicity testing using mice model. During immune magnetic polymerase chain reaction IM-PCR protocol, aerolysin specific antibodies were bound with immuno magnetic binding. Sensitivity and specificity tests for IM-PCR were conducted. RESULTS: There was high detection of Aeromonas using IM-PCR (12.4 %) technique when compared to low isolation with culture (5.1%). Our study confirmed that some strains of enterotoxic Aeromonas strains were uncultivable. Enterotoxicity tests on culture isolates revealed many strains were negative. IM-PCR detected high, (62/500) rate of identification of Aeromonas with aerolysin toxin gene. Aeromonas species identified after IM-PCR were A. hydrophila (40.3% ), A. veronii (17.7 %), A. caviae (14.5 %), A. trota (11.2 %), A. jandei (9.6 %) and A. schuberti (6.4%). All A. trota strains were undetected by cultivation. CONCLUSION: High sensitivity and specificity of IM-PCR are due to preparation of aerolysin antibodies and immuno magnetic binding, prior to PCR. Since diseases due to Aeromonas are increasingly reported, IM-PCR is recommended for detection from clinical specimens. Makerere Medical School 2019-06 /pmc/articles/PMC6794535/ /pubmed/31656487 http://dx.doi.org/10.4314/ahs.v19i2.27 Text en © 2019 Subbaram et al. Licensee African Health Sciences. This is an Open Access article distributed under the terms of the Creative commons Attribution License (https://creativecommons.org/licenses/BY/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Articles
Subbaram, Kannan
Gatasheh, Mansour K
Al Azzam, Khaldun M
Kannan, Hemalatha
Molecular identification of diarrheal Aeromonas using immuno magnetic polymerase chain reaction (IM-PCR) technique: a comparative study with conventional culture method
title Molecular identification of diarrheal Aeromonas using immuno magnetic polymerase chain reaction (IM-PCR) technique: a comparative study with conventional culture method
title_full Molecular identification of diarrheal Aeromonas using immuno magnetic polymerase chain reaction (IM-PCR) technique: a comparative study with conventional culture method
title_fullStr Molecular identification of diarrheal Aeromonas using immuno magnetic polymerase chain reaction (IM-PCR) technique: a comparative study with conventional culture method
title_full_unstemmed Molecular identification of diarrheal Aeromonas using immuno magnetic polymerase chain reaction (IM-PCR) technique: a comparative study with conventional culture method
title_short Molecular identification of diarrheal Aeromonas using immuno magnetic polymerase chain reaction (IM-PCR) technique: a comparative study with conventional culture method
title_sort molecular identification of diarrheal aeromonas using immuno magnetic polymerase chain reaction (im-pcr) technique: a comparative study with conventional culture method
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6794535/
https://www.ncbi.nlm.nih.gov/pubmed/31656487
http://dx.doi.org/10.4314/ahs.v19i2.27
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