Development and Evaluation of Duplex MIRA-qPCR Assay for Simultaneous Detection of Staphylococcus aureus and non-aureus Staphylococci Article Swipe
Jiulian Lai
,
Zhenzhou Huang
,
Yue Xiao
,
Keyi Yu
,
Xuemei Bai
,
He Gao
,
Hang Dai
,
Xiaoning Liu
,
Duochun Wang
·
YOU?
·
· 2022
· Open Access
·
· DOI: https://doi.org/10.3390/microorganisms10091734
YOU?
·
· 2022
· Open Access
·
· DOI: https://doi.org/10.3390/microorganisms10091734
Staphylococcus spp., especially Staphylococcus aureus (S. aureus), is an important pathogen in hospital-acquired infection and food poisoning. Here, we developed a multienzyme isothermal rapid amplification combined with duplex quantitative PCR (duplex MIRA-qPCR) method, which can simultaneously detect the S. aureus species-specific conserved gene FMN-bgsfp and the Staphylococcus genus-specific conserved gene tuf. This assay enabled the amplification of DNA within 20 min at a constant temperature of 39 °C. Specificity analysis indicated that all nine common Staphylococcus species were positive and non-Staphylococcus spp. were negative for tuf gene, whereas S. aureus was positive, non-aureus Staphylococci species and non-Staphylococcus spp. were negative for FMN-bgsfp gene, suggesting that duplex MIRA-qPCR exhibited high specificity. Meanwhile, the sensitivity was tested and the limit of detection (LoD) was 3 × 102 CFU/mL. The coefficient variation values ranged from 0.13% to 2.09%, indicating that the assay had good repeatability. Furthermore, all the nine common Staphylococcus species (including S. aureus) could be detected from four kinds of simulated samples and the LoD of S. aureus was 8.56 × 103 CFU/mL. In conclusion, the duplex MIRA-qPCR has advantages of stronger specificity, lower detection threshold, shorter detection time, and simpler operation, which is an effective tool to detect S. aureus and non-aureus Staphylococci spp. infections rapidly.
Related Topics
Concepts
Staphylococcus aureus
Biology
Microbiology
Duplex (building)
Staphylococcus
Gene
DNA
Genetics
Bacteria
Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.3390/microorganisms10091734
- https://www.mdpi.com/2076-2607/10/9/1734/pdf?version=1661924500
- OA Status
- gold
- Cited By
- 16
- References
- 67
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4293761287
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4293761287Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.3390/microorganisms10091734Digital Object Identifier
- Title
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Development and Evaluation of Duplex MIRA-qPCR Assay for Simultaneous Detection of Staphylococcus aureus and non-aureus StaphylococciWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
- Publication year
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2022Year of publication
- Publication date
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2022-08-27Full publication date if available
- Authors
-
Jiulian Lai, Zhenzhou Huang, Yue Xiao, Keyi Yu, Xuemei Bai, He Gao, Hang Dai, Xiaoning Liu, Duochun WangList of authors in order
- Landing page
-
https://doi.org/10.3390/microorganisms10091734Publisher landing page
- PDF URL
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https://www.mdpi.com/2076-2607/10/9/1734/pdf?version=1661924500Direct link to full text PDF
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YesWhether a free full text is available
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goldOpen access status per OpenAlex
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https://www.mdpi.com/2076-2607/10/9/1734/pdf?version=1661924500Direct OA link when available
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Staphylococcus aureus, Biology, Microbiology, Duplex (building), Staphylococcus, Gene, DNA, Genetics, BacteriaTop concepts (fields/topics) attached by OpenAlex
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16Total citation count in OpenAlex
- Citations by year (recent)
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2025: 5, 2024: 7, 2023: 4Per-year citation counts (last 5 years)
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67Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.at | 61 |
| abstract_inverted_index.be | 153 |
| abstract_inverted_index.in | 11 |
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| abstract_inverted_index.of | 56, 65, 118, 158, 164, 179 |
| abstract_inverted_index.to | 133, 196 |
| abstract_inverted_index.we | 18 |
| abstract_inverted_index.× | 123, 169 |
| abstract_inverted_index.(S. | 5 |
| abstract_inverted_index.102 | 124 |
| abstract_inverted_index.103 | 170 |
| abstract_inverted_index.DNA | 57 |
| abstract_inverted_index.LoD | 163 |
| abstract_inverted_index.PCR | 29 |
| abstract_inverted_index.The | 126 |
| abstract_inverted_index.all | 72, 143 |
| abstract_inverted_index.and | 14, 44, 79, 95, 115, 161, 188, 200 |
| abstract_inverted_index.can | 34 |
| abstract_inverted_index.for | 84, 100 |
| abstract_inverted_index.had | 139 |
| abstract_inverted_index.has | 177 |
| abstract_inverted_index.min | 60 |
| abstract_inverted_index.the | 37, 45, 54, 111, 116, 137, 144, 162, 174 |
| abstract_inverted_index.tuf | 85 |
| abstract_inverted_index.was | 90, 113, 121, 167 |
| abstract_inverted_index.8.56 | 168 |
| abstract_inverted_index.This | 51 |
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| abstract_inverted_index.four | 156 |
| abstract_inverted_index.from | 131, 155 |
| abstract_inverted_index.gene | 42, 49 |
| abstract_inverted_index.good | 140 |
| abstract_inverted_index.high | 108 |
| abstract_inverted_index.nine | 73, 145 |
| abstract_inverted_index.spp. | 81, 97, 203 |
| abstract_inverted_index.that | 71, 104, 136 |
| abstract_inverted_index.tool | 195 |
| abstract_inverted_index.tuf. | 50 |
| abstract_inverted_index.were | 77, 82, 98 |
| abstract_inverted_index.with | 26 |
| abstract_inverted_index.°C. | 67 |
| abstract_inverted_index.(LoD) | 120 |
| abstract_inverted_index.0.13% | 132 |
| abstract_inverted_index.Here, | 17 |
| abstract_inverted_index.assay | 52, 138 |
| abstract_inverted_index.could | 152 |
| abstract_inverted_index.gene, | 86, 102 |
| abstract_inverted_index.kinds | 157 |
| abstract_inverted_index.limit | 117 |
| abstract_inverted_index.lower | 182 |
| abstract_inverted_index.rapid | 23 |
| abstract_inverted_index.spp., | 1 |
| abstract_inverted_index.time, | 187 |
| abstract_inverted_index.which | 33, 191 |
| abstract_inverted_index.2.09%, | 134 |
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| abstract_inverted_index.common | 74, 146 |
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| abstract_inverted_index.duplex | 27, 105, 175 |
| abstract_inverted_index.ranged | 130 |
| abstract_inverted_index.tested | 114 |
| abstract_inverted_index.values | 129 |
| abstract_inverted_index.within | 58 |
| abstract_inverted_index.(duplex | 30 |
| abstract_inverted_index.CFU/mL. | 125, 171 |
| abstract_inverted_index.aureus) | 151 |
| abstract_inverted_index.enabled | 53 |
| abstract_inverted_index.method, | 32 |
| abstract_inverted_index.samples | 160 |
| abstract_inverted_index.shorter | 185 |
| abstract_inverted_index.simpler | 189 |
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| abstract_inverted_index.whereas | 87 |
| abstract_inverted_index.analysis | 69 |
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| abstract_inverted_index.detected | 154 |
| abstract_inverted_index.negative | 83, 99 |
| abstract_inverted_index.pathogen | 10 |
| abstract_inverted_index.positive | 78 |
| abstract_inverted_index.rapidly. | 205 |
| abstract_inverted_index.stronger | 180 |
| abstract_inverted_index.FMN-bgsfp | 43, 101 |
| abstract_inverted_index.MIRA-qPCR | 106, 176 |
| abstract_inverted_index.conserved | 41, 48 |
| abstract_inverted_index.detection | 119, 183, 186 |
| abstract_inverted_index.developed | 19 |
| abstract_inverted_index.effective | 194 |
| abstract_inverted_index.exhibited | 107 |
| abstract_inverted_index.important | 9 |
| abstract_inverted_index.indicated | 70 |
| abstract_inverted_index.infection | 13 |
| abstract_inverted_index.positive, | 91 |
| abstract_inverted_index.simulated | 159 |
| abstract_inverted_index.variation | 128 |
| abstract_inverted_index.(including | 149 |
| abstract_inverted_index.MIRA-qPCR) | 31 |
| abstract_inverted_index.Meanwhile, | 110 |
| abstract_inverted_index.advantages | 178 |
| abstract_inverted_index.especially | 2 |
| abstract_inverted_index.indicating | 135 |
| abstract_inverted_index.infections | 204 |
| abstract_inverted_index.isothermal | 22 |
| abstract_inverted_index.non-aureus | 92, 201 |
| abstract_inverted_index.operation, | 190 |
| abstract_inverted_index.poisoning. | 16 |
| abstract_inverted_index.suggesting | 103 |
| abstract_inverted_index.threshold, | 184 |
| abstract_inverted_index.Specificity | 68 |
| abstract_inverted_index.coefficient | 127 |
| abstract_inverted_index.conclusion, | 173 |
| abstract_inverted_index.multienzyme | 21 |
| abstract_inverted_index.sensitivity | 112 |
| abstract_inverted_index.temperature | 64 |
| abstract_inverted_index.Furthermore, | 142 |
| abstract_inverted_index.quantitative | 28 |
| abstract_inverted_index.specificity, | 181 |
| abstract_inverted_index.specificity. | 109 |
| abstract_inverted_index.Staphylococci | 93, 202 |
| abstract_inverted_index.amplification | 24, 55 |
| abstract_inverted_index.Staphylococcus | 0, 3, 46, 75, 147 |
| abstract_inverted_index.genus-specific | 47 |
| abstract_inverted_index.repeatability. | 141 |
| abstract_inverted_index.simultaneously | 35 |
| abstract_inverted_index.species-specific | 40 |
| abstract_inverted_index.hospital-acquired | 12 |
| abstract_inverted_index.non-Staphylococcus | 80, 96 |
| cited_by_percentile_year.max | 99 |
| cited_by_percentile_year.min | 97 |
| corresponding_author_ids | https://openalex.org/A5080900897, https://openalex.org/A5024125020 |
| countries_distinct_count | 1 |
| institutions_distinct_count | 9 |
| corresponding_institution_ids | https://openalex.org/I184490438, https://openalex.org/I4210146685, https://openalex.org/I4391768093, https://openalex.org/I76214153 |
| sustainable_development_goals[0].id | https://metadata.un.org/sdg/2 |
| sustainable_development_goals[0].score | 0.44999998807907104 |
| sustainable_development_goals[0].display_name | Zero hunger |
| citation_normalized_percentile.value | 0.78911798 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | False |