Chaemsaithong, Piya, Romero, Roberto, Pongchaikul, Pisut 
ORCID: 0000-0001-8757-3554, Vivithanaporn, Pornpun, Lertrut, Waranyu, Jaovisidha, Adithep, Mongkolsuk, Paninee, Nitayanon, Perapon, Pongsuktavorn, Khontawan, Kamlungkuea, Threebhorn et al (show 6 more authors)
(2023)
Rapid diagnosis of intra-amniotic infection using nanopore-based sequencing.
JOURNAL OF PERINATAL MEDICINE, 51 (6).
pp. 769-774.
Abstract
<h4>Objectives</h4>Early diagnosis and treatment of intra-amniotic infection is crucial. Rapid pathogen identification allows for a definite diagnosis and enables proper management. We determined whether the 16S amplicon sequencing performed by a nanopore sequencing technique make possible rapid bacterial identification at the species level in intra-amniotic infection.<h4>Methods</h4>Five cases of confirmed intra-amniotic infection, determined by either cultivation or 16S rDNA polymerase chain reaction (PCR) Sanger sequencing, and 10 cases of women who underwent mid-trimester genetic amniocentesis were included. DNA was extracted from amniotic fluid and PCR was performed on the full-length 16S rDNA. Nanopore sequencing was performed. The results derived from nanopore sequencing were compared with those derived from cultivation and Sanger sequencing methods.<h4>Results</h4>Bacteria were successfully detected from amniotic fluid using nanopore sequencing in all cases of intra-amniotic infection. Nanopore sequencing identified additional bacterial species and polymicrobial infections. All patients who underwent a mid-trimester amniocentesis had negative cultures, negative 16S PCR Sanger sequencing and nanopore sequencing. Identification of the microorganisms using nanopore sequencing technique at the bacterial species level was achieved within 5-9 h from DNA extraction.<h4>Conclusions</h4>This is the first study demonstrating that the nanopore sequencing technique is capable of rapid diagnosis of intra-amniotic infection using fresh amniotic fluid samples.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | 16S rDNA, amniocentesis, amniotic fluid, bacteria, infection, intra-amniotic inflammation, long-read sequencing, microorganism, preterm labor, Sanger sequencing |
| Divisions: | Faculty of Health and Life Sciences Faculty of Health and Life Sciences > Institute of Infection, Veterinary and Ecological Sciences |
| Depositing User: | Symplectic Admin |
| Date Deposited: | 28 Feb 2024 10:32 |
| Last Modified: | 28 Feb 2024 10:33 |
| DOI: | 10.1515/jpm-2022-0504 |
| Open Access URL: | https://doi.org/10.1515/jpm-2022-0504 |
| Related URLs: | |
| URI: | https://livrepository.liverpool.ac.uk/id/eprint/3178947 |
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