High Resolution Melt Assays to Detect and Identify <i>Vibrio parahaemolyticus</i>, <i>Bacillus cereus</i>, <i>Escherichia coli</i>, and <i>Clostridioides difficile</i> Bacteria
Over one hundred bacterial species have been determined to comprise the human microbiota in a healthy individual. Bacteria including <i>Escherichia coli</i>, <i>Bacillus cereus</i>, <i>Clostridioides difficile</i>, and <i>Vibrio parahaemolyticus</i> ar...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-04-01
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| Series: | Microorganisms |
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| Online Access: | https://www.mdpi.com/2076-2607/8/4/561 |
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| author | Allison C. Bender Jessica A. Faulkner Katherine Tulimieri Thomas H. Boise Kelly M. Elkins |
| author_facet | Allison C. Bender Jessica A. Faulkner Katherine Tulimieri Thomas H. Boise Kelly M. Elkins |
| author_sort | Allison C. Bender |
| collection | DOAJ |
| description | Over one hundred bacterial species have been determined to comprise the human microbiota in a healthy individual. Bacteria including <i>Escherichia coli</i>, <i>Bacillus cereus</i>, <i>Clostridioides difficile</i>, and <i>Vibrio parahaemolyticus</i> are found inside of the human body and <i>B. cereus</i> and <i>E. coli</i> are also found on the skin. These bacteria can act as human pathogens upon ingestion of contaminated food or water, if they enter an open wound, or antibiotics, and environment or stress can alter the microbiome. In this study, we present new polymerase chain reaction (PCR) high-resolution melt (HRM) assays to detect and identify the above microorganisms. Amplified DNA from <i>C. difficile</i>, <i>E. coli, B. cereus</i>, and <i>V. parahaemolyticus</i> melted at 80.37 ± 0.45 °C, 82.15 ± 0.37 °C, 84.43 ± 0.50 °C, and 86.74 ± 0.65 °C, respectively. A triplex PCR assay was developed to simultaneously detect and identify <i>E. coli</i>, <i>B. cereus</i>, and <i>V. parahaemolyticus</i>, and cultured microorganisms were successfully amplified, detected, and identified. The assays demonstrated sensitivity, specificity, reproducibility, and robustness in testing. |
| first_indexed | 2024-03-10T20:29:25Z |
| format | Article |
| id | doaj.art-d1fc2fff998a4f94ba41b2bc34e5f2ab |
| institution | Directory Open Access Journal |
| issn | 2076-2607 |
| language | English |
| last_indexed | 2024-03-10T20:29:25Z |
| publishDate | 2020-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Microorganisms |
| spelling | doaj.art-d1fc2fff998a4f94ba41b2bc34e5f2ab2023-11-19T21:32:37ZengMDPI AGMicroorganisms2076-26072020-04-018456110.3390/microorganisms8040561High Resolution Melt Assays to Detect and Identify <i>Vibrio parahaemolyticus</i>, <i>Bacillus cereus</i>, <i>Escherichia coli</i>, and <i>Clostridioides difficile</i> BacteriaAllison C. Bender0Jessica A. Faulkner1Katherine Tulimieri2Thomas H. Boise3Kelly M. Elkins4Chemistry Department, Towson University, 8000 York Road, Towson, MD 21252, USAChemistry Department, Towson University, 8000 York Road, Towson, MD 21252, USAChemistry Department, Towson University, 8000 York Road, Towson, MD 21252, USAChemistry Department, Towson University, 8000 York Road, Towson, MD 21252, USAChemistry Department, Towson University, 8000 York Road, Towson, MD 21252, USAOver one hundred bacterial species have been determined to comprise the human microbiota in a healthy individual. Bacteria including <i>Escherichia coli</i>, <i>Bacillus cereus</i>, <i>Clostridioides difficile</i>, and <i>Vibrio parahaemolyticus</i> are found inside of the human body and <i>B. cereus</i> and <i>E. coli</i> are also found on the skin. These bacteria can act as human pathogens upon ingestion of contaminated food or water, if they enter an open wound, or antibiotics, and environment or stress can alter the microbiome. In this study, we present new polymerase chain reaction (PCR) high-resolution melt (HRM) assays to detect and identify the above microorganisms. Amplified DNA from <i>C. difficile</i>, <i>E. coli, B. cereus</i>, and <i>V. parahaemolyticus</i> melted at 80.37 ± 0.45 °C, 82.15 ± 0.37 °C, 84.43 ± 0.50 °C, and 86.74 ± 0.65 °C, respectively. A triplex PCR assay was developed to simultaneously detect and identify <i>E. coli</i>, <i>B. cereus</i>, and <i>V. parahaemolyticus</i>, and cultured microorganisms were successfully amplified, detected, and identified. The assays demonstrated sensitivity, specificity, reproducibility, and robustness in testing.https://www.mdpi.com/2076-2607/8/4/561dysbiosispathogen<i>Escherichia coli</i><i>Vibrio parahaemolyticus</i><i>Clostridioides difficile</i><i>Bacillus cereus</i> |
| spellingShingle | Allison C. Bender Jessica A. Faulkner Katherine Tulimieri Thomas H. Boise Kelly M. Elkins High Resolution Melt Assays to Detect and Identify <i>Vibrio parahaemolyticus</i>, <i>Bacillus cereus</i>, <i>Escherichia coli</i>, and <i>Clostridioides difficile</i> Bacteria Microorganisms dysbiosis pathogen <i>Escherichia coli</i> <i>Vibrio parahaemolyticus</i> <i>Clostridioides difficile</i> <i>Bacillus cereus</i> |
| title | High Resolution Melt Assays to Detect and Identify <i>Vibrio parahaemolyticus</i>, <i>Bacillus cereus</i>, <i>Escherichia coli</i>, and <i>Clostridioides difficile</i> Bacteria |
| title_full | High Resolution Melt Assays to Detect and Identify <i>Vibrio parahaemolyticus</i>, <i>Bacillus cereus</i>, <i>Escherichia coli</i>, and <i>Clostridioides difficile</i> Bacteria |
| title_fullStr | High Resolution Melt Assays to Detect and Identify <i>Vibrio parahaemolyticus</i>, <i>Bacillus cereus</i>, <i>Escherichia coli</i>, and <i>Clostridioides difficile</i> Bacteria |
| title_full_unstemmed | High Resolution Melt Assays to Detect and Identify <i>Vibrio parahaemolyticus</i>, <i>Bacillus cereus</i>, <i>Escherichia coli</i>, and <i>Clostridioides difficile</i> Bacteria |
| title_short | High Resolution Melt Assays to Detect and Identify <i>Vibrio parahaemolyticus</i>, <i>Bacillus cereus</i>, <i>Escherichia coli</i>, and <i>Clostridioides difficile</i> Bacteria |
| title_sort | high resolution melt assays to detect and identify i vibrio parahaemolyticus i i bacillus cereus i i escherichia coli i and i clostridioides difficile i bacteria |
| topic | dysbiosis pathogen <i>Escherichia coli</i> <i>Vibrio parahaemolyticus</i> <i>Clostridioides difficile</i> <i>Bacillus cereus</i> |
| url | https://www.mdpi.com/2076-2607/8/4/561 |
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