Multiplex PCR assay to identify clinically important Aeromonas species
ABSTRACT The genus Aeromonas is increasingly implicated in human infections. However, accurate species-level identification remains challenging, particularly in clinical microbiology laboratories. This study aimed to develop a multiplex polymerase chain reaction (PCR) assay to identify four Aeromona...
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| Format: | Article |
| Language: | English |
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American Society for Microbiology
2025-05-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.03331-24 |
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| author | Aki Sakurai Naoto Hosokawa Daisuke Ohkushi Sohei Harada Yasufumi Matsumura Naoya Itoh Kazuhiro Ishikawa Sho Saito Takayuki Sakurai Ryota Hase Takehiro Hashimoto Yohei Doi Masahiro Suzuki |
| author_facet | Aki Sakurai Naoto Hosokawa Daisuke Ohkushi Sohei Harada Yasufumi Matsumura Naoya Itoh Kazuhiro Ishikawa Sho Saito Takayuki Sakurai Ryota Hase Takehiro Hashimoto Yohei Doi Masahiro Suzuki |
| author_sort | Aki Sakurai |
| collection | DOAJ |
| description | ABSTRACT The genus Aeromonas is increasingly implicated in human infections. However, accurate species-level identification remains challenging, particularly in clinical microbiology laboratories. This study aimed to develop a multiplex polymerase chain reaction (PCR) assay to identify four Aeromonas species—Aeromonas hydrophila, Aeromonas caviae, Aeromonas veronii, and Aeromonas dhakensis—most frequently associated with human infectious diseases. A total of 788 whole genome sequencing (WGS) data sets from 31 Aeromonas species were analyzed to identify open reading frames (ORFs) specifically present in A. hydrophila, A. caviae, A. veronii, and A. dhakensis. Primer sets were designed based on sequences of ORFs specific to each species to develop a multiplex PCR assay. To validate the efficacy of the assay, 256 clinical Aeromonas isolates were tested, and the results were compared with taxonomic affiliation inferred by WGS data, along with 19 type strains. The multiplex PCR successfully identified all strains of the four target species and produced no amplification in non-target species strains except the band for internal control. The multiplex PCR enables rapid and reliable identification of four Aeromonas spp. commonly involved in human infectious diseases.IMPORTANCEThe multiplex PCR assay facilitates accurate identification of clinically important Aeromonas spp. in clinical microbiology laboratories, providing crucial information to guide appropriate antimicrobial therapy and advance understanding of the epidemiology of Aeromonas spp. |
| format | Article |
| id | doaj-art-3b8ecf72feb342de990347feea7d33d5 |
| institution | Kabale University |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | Microbiology Spectrum |
| spelling | doaj-art-3b8ecf72feb342de990347feea7d33d52025-08-20T03:52:10ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-05-0113510.1128/spectrum.03331-24Multiplex PCR assay to identify clinically important Aeromonas speciesAki Sakurai0Naoto Hosokawa1Daisuke Ohkushi2Sohei Harada3Yasufumi Matsumura4Naoya Itoh5Kazuhiro Ishikawa6Sho Saito7Takayuki Sakurai8Ryota Hase9Takehiro Hashimoto10Yohei Doi11Masahiro Suzuki12Department of Infectious Diseases, Fujita Health University School of Medicine, Aichi, JapanDepartment of Infectious Diseases, Kameda Medical Center, Chiba, JapanDepartment of Infectious Diseases, Cancer Institute Hospital, Japanese Foundation for Cancer Research, Tokyo, JapanDepartment of Microbiology and Infectious Diseases, Toho University School of Medicine, Tokyo, JapanDepartment of Clinical Laboratory Medicine, Kyoto University Graduate School of Medicine, Kyoto, JapanDepartment of Infectious Diseases, Graduate School of Medical Sciences, Nagoya City University, Aichi, JapanDepartment of Infectious Diseases, St. Luke’s International Hospital, Tokyo, JapanDisease Control and Prevention Center, National Center for Global Health and Medicine, Tokyo, JapanDepartment of Infectious Diseases, NTT Medical Center, Tokyo, JapanDepartment of Infectious Diseases, Japanese Red Cross Narita Hospital, Chiba, JapanHospital Infection Control Center, Oita University Hospital, Oita, JapanDepartment of Infectious Diseases, Fujita Health University School of Medicine, Aichi, JapanDepartment of Microbiology, Fujita Health University School of Medicine, Aichi, JapanABSTRACT The genus Aeromonas is increasingly implicated in human infections. However, accurate species-level identification remains challenging, particularly in clinical microbiology laboratories. This study aimed to develop a multiplex polymerase chain reaction (PCR) assay to identify four Aeromonas species—Aeromonas hydrophila, Aeromonas caviae, Aeromonas veronii, and Aeromonas dhakensis—most frequently associated with human infectious diseases. A total of 788 whole genome sequencing (WGS) data sets from 31 Aeromonas species were analyzed to identify open reading frames (ORFs) specifically present in A. hydrophila, A. caviae, A. veronii, and A. dhakensis. Primer sets were designed based on sequences of ORFs specific to each species to develop a multiplex PCR assay. To validate the efficacy of the assay, 256 clinical Aeromonas isolates were tested, and the results were compared with taxonomic affiliation inferred by WGS data, along with 19 type strains. The multiplex PCR successfully identified all strains of the four target species and produced no amplification in non-target species strains except the band for internal control. The multiplex PCR enables rapid and reliable identification of four Aeromonas spp. commonly involved in human infectious diseases.IMPORTANCEThe multiplex PCR assay facilitates accurate identification of clinically important Aeromonas spp. in clinical microbiology laboratories, providing crucial information to guide appropriate antimicrobial therapy and advance understanding of the epidemiology of Aeromonas spp.https://journals.asm.org/doi/10.1128/spectrum.03331-24Aeromonasmultiplex polymerase chain reactionopen reading framespecies identificationrapid identification techniquehuman infections |
| spellingShingle | Aki Sakurai Naoto Hosokawa Daisuke Ohkushi Sohei Harada Yasufumi Matsumura Naoya Itoh Kazuhiro Ishikawa Sho Saito Takayuki Sakurai Ryota Hase Takehiro Hashimoto Yohei Doi Masahiro Suzuki Multiplex PCR assay to identify clinically important Aeromonas species Microbiology Spectrum Aeromonas multiplex polymerase chain reaction open reading frame species identification rapid identification technique human infections |
| title | Multiplex PCR assay to identify clinically important Aeromonas species |
| title_full | Multiplex PCR assay to identify clinically important Aeromonas species |
| title_fullStr | Multiplex PCR assay to identify clinically important Aeromonas species |
| title_full_unstemmed | Multiplex PCR assay to identify clinically important Aeromonas species |
| title_short | Multiplex PCR assay to identify clinically important Aeromonas species |
| title_sort | multiplex pcr assay to identify clinically important aeromonas species |
| topic | Aeromonas multiplex polymerase chain reaction open reading frame species identification rapid identification technique human infections |
| url | https://journals.asm.org/doi/10.1128/spectrum.03331-24 |
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