Profiling system of oral microbiota utilizing polymerase chain reaction-restriction fragment length polymorphism analysis

Sano, Hiroto; Wakui, Anna; Kawachi, Miho; Washio, Jumpei; Abiko, Yoshihiro; Mayanagi, Gen; Yamaki, Kouya; Tanaka, Kaori; Takahashi, Nobuhiro; Sato, Takuichi

doi:10.1016/j.job.2021.05.003

Cited by 11 publications

(3 citation statements)

References 15 publications

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“…For further identification, two single colonies from the Columbia blood agar were selected. After genomic DNA extraction, the general primers [ 11 ] (27 F and 1492R, Table 1 ) for 16S rDNA were used for PCR amplification. Following gel electrophoresis, purification, recovery, cloning and sequencing, the sequencing results (GenBank accession numbers MZ220349 and MZ220350) were analyzed by BLAST, and the homology to the Olsenella uli sequence in the GenBank was 99%, and the NCBI number referenced was CP002106.1.…”

Section: Case Presentationmentioning

confidence: 99%

Olsenella uli-induced pneumonia: a case report

Yan

et al. 2022

Ann Clin Microbiol Antimicrob

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Background Olsenella uli is anaerobic or microaerophilic bacteria, commonly found in oral cavity or gastrointestinal tract, which has not been reported to be associated with lower respiratory tract infection. Herein, we report the first case of Olsenella uli infection in the lung. Case presentation A 70-year-old male farmer with no history of other respiratory tract diseases developed a cough with bloody sputum three times a day without obvious causes or other concomitant symptoms. After a period of treatment with empirical antibiotic, his condition did not improve. The computed tomography (CT) and lung biopsy results indicated bilateral pneumonia, and Olsenella uli was identified by micromorphology, sequence analysis and mass spectrometry analysis recovered from sputum. Ceftazidime, a third generation cephalosporin was used for the treatment, and the patient recovered after 10 days. Conclusions Our report suggests a causative role of gingival bacteria in the pathogenesis of pneumonia, thus early diagnosis and prompt antibiotic therapy may play a role in the treatment of Olsenella uli induced pneumonia.

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Section: Case Presentationmentioning

confidence: 99%

Olsenella uli-induced pneumonia: a case report

Yan

et al. 2022

Ann Clin Microbiol Antimicrob

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“…Samples with higher concentrations and better quality of DNA were amplified using the PCR technique in a thermocycler (Bio-Rad T100™ thermocycler, Hercules, CA, USA), using the universal primers 27F, 5′-AGAGTTTGATCMTGGCTCAG-3′ and 1492R, 5′-TACGGYTACCTTGTTACGACTT-3′ (Sigma-Aldrich, Darmstadt, Germany, part number 200-00485), which amplify the 16S rRNA gene [ 13 ]. The reaction was prepared in a final volume of 50 µL, which contained 25 µL of DreamTaq Hot Start PCR Master Mix (2X) (DreamTaq™ Hot Start PCR Master Mix, Thermo Fisher Scientific, part number K9011), 1 µL of each 10 mM primer, 2 µL of DNA (~157 ng/µL) and sufficient molecular grade water to reach the desired volume.…”

Section: Methodsmentioning

confidence: 99%

A New and Profitable Protocol to DNA Extraction in Limnospira maxima

Pineda-Rodriguez

Pompelli

Jarma‐Orozco

et al. 2023

MPs

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Limnospira maxima is a remarkable organism showing great potential as a versatile and sustainable food source, offering a powerful solution to address the pressing issues of malnutrition and undernourishment worldwide. L. maxima contains high amounts of proteins, vitamins, minerals, and essential fatty acids. It can be grown in both bioreactors and open systems; however, before considering industrial production, optimization studies of the cultivation must be conducted to obtain knowledge about the ideal environmental conditions. Additionally, for the molecular typing of L. maxima strains and their industrial scaling, high-quality and large quantity DNA extraction is required. Notwithstanding, DNA extraction from L. maxima can be challenging due to the low amount of DNA in cells and the presence of difficult-to-remove substances such as polysaccharides and polyphenols. In this study, the quality and quantity of DNA extracted from two types of L. maxima samples (Limnospira maxima strain SISCA accession GenBank: OR195505.1) were evaluated using three commercially available DNA extraction kits and two types of input biological material. The results showed that Pbact-P kit had the highest quantity and quality of DNA, while CTAB-P allowed for a higher quantity and quality of RNA, making them optimal protocols for nucleic acid extraction to improve PCR, rt-PCR, and genome sequencing of L. maxima compared with other extraction methods.

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“…For further identi cation, two single colonies were selected from Columbia blood AGAR. After genomic DNA extraction, PCR ampli cation was performed using universal primers [12] (27F and 1492R, Table 1) targeting 16S rDNA. Next, gel electrophoresis, puri cation, recovery, cloning, and sequencing were conducted.…”

Section: Case Presentationmentioning

confidence: 99%

Lautropia mirabilis-induced peritoneal dialysis-associated peritonitis: a case report

Liu

Yan

et al. 2023

Preprint

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Background Lautropia mirabilis is a Gram-negative bacterium mainly isolated from the mouth and upper respiratory tract. To date, only two cases of peritoneal dialysis-associated peritonitis induced by Lautropia mirabilis have been globally reported: one in Australia and one in Portugal, but Lautropia mirabilis has not been cultured. Here, we report the third case of peritoneal dialysis-associated peritonitis caused by Lautropia mirabilis in the world. Furthermore, we cultivated this bacterium to improve the existing knowledge on the etiology of this condition and provide clinical guidance for its treatment. Case presentation: A 49-year-old female with renal insufficiency for more than five years was admitted to our hospital. She had been on peritoneal dialysis for six months. The patient had diarrhea and turbid peritoneal dialysis fluid for three days. With indication of infection by routine results of peritoneal dialysis fluid, mass spectrometer identification and gene sequencing of peritoneal dialysis fluid culture were subjected and revealed infection with Lautropia mirabilis. After anti-infection treatment with gentamicin and ceftazidime, the patient recovered. Conclusions Our findings suggest that Lautropia mirabilis plays an important role in the pathogenesis of peritoneal dialysis-associated peritonitis. Early diagnosis and timely antibiotic treatment may be essential for the treatment of this infection.

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Profiling system of oral microbiota utilizing polymerase chain reaction-restriction fragment length polymorphism analysis

Cited by 11 publications

References 15 publications

Olsenella uli-induced pneumonia: a case report

Olsenella uli-induced pneumonia: a case report

A New and Profitable Protocol to DNA Extraction in Limnospira maxima

Lautropia mirabilis-induced peritoneal dialysis-associated peritonitis: a case report

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