AIMS MicrobiologyPub Date : 2022-09-15eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022024
T Amrouche, M L Chikindas
{"title":"Probiotics for immunomodulation in prevention against respiratory viral infections with special emphasis on COVID-19.","authors":"T Amrouche, M L Chikindas","doi":"10.3934/microbiol.2022024","DOIUrl":"https://doi.org/10.3934/microbiol.2022024","url":null,"abstract":"<p><p>COVID-19 pandemic is currently causing high mortality and economic crisis, and several drugs-based therapeutic strategies and vaccines are unfortunately used with little efficiency. Therefore, here is an urgent need to provide additives therapies that prevent or improve symptoms in populations infected by SARS-CoV-2 variants. This review aimed to examine relevant scientific information related to SARS-CoV-2 and host antiviral immunity, as well the possible role of probiotics in gut-lung cross talk pathways to promote lung immune response to COVID-19 infection. We searched online databases such as PubMed, Embase, Chinese databases, and selected articles and studies with relevant data reported on COVID-19 and other respiratory viral infections. Recent research highlighted potential immunomodulatory activities of probiotics assessed in animal models and clinical trials. However, the role of probiotics and gut microbiome in COVID-19 management, and approaches with significant understanding in molecular mechanism of probiotic action remain poorly investigated. Clinical investigations as well as animal model studies published have demonstrated that probiotics such as <i>Lactobacillus rhamnosus</i> and <i>Bifidumbacterium lactis</i> HN019, may influence positively not only microbiota balance but also antiviral immunity by improving both innate and adaptive responses and controlling inflammatory reaction in respiratory viral infection. Given the immunological interactions in gut-lung axis and the crucial role of probiotics in modulating immune responses by promoting dendritic cells (DCs) to regulate T cell responses, we hypothesized that application of probiotics may be successful in prevention or treatment of both intestinal disorders and airway diseases in patients with COVID-19.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 3","pages":"338-356"},"PeriodicalIF":4.8,"publicationDate":"2022-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9576497/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40657934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"Potential use of <i>Bacillus paramycoides</i> for the production of the biopolymer polyhydroxybutyrate from leftover carob fruit agro-waste.","authors":"Leila Djerrab, Zohra Chekroud, Amer Rouabhia, Mohamed Abdesselem Dems, Imane Attailia, Luis Isidoro Romero Garcia, Mustapha Adnane Smadi","doi":"10.3934/microbiol.2022023","DOIUrl":"https://doi.org/10.3934/microbiol.2022023","url":null,"abstract":"<p><p>This study was designed to investigate, at a laboratory scale, the possibility of valorizing the leftover carob fruits to produce the eco-friendly biopolymer polyhydroxybutyrate (PHB) by using the bacterial strain <i>Bacillus paramycoides</i>, which has been isolated from the botanical garden of Skikda University in Algeria. The PHB production was tested under various conditions: a pH of 3-8, temperature range of 30-44 °C, carob extracted molasses concentration of 2-8% v/v, an incubation time of 24-96 h and an agitation speed of 150-300 rpm. The effects of different nitrogen sources and carob extracted molasses treatment types were also investigated. The PHB concentration was determined quantitatively as crotonic acid by measuring the absorbance at 300 nm. Cell growth was quantified by measuring the density of the culture at 600 nm. The presence of PHB was confirmed by applying high-performance liquid chromatography (HPLC) using an Aminex HPX-87H and implementing gas chromatography analysis. The best yield of PHB synthesis was obtained by using 6% v/v of 5 M H<sub>2</sub>SO<sub>4</sub> treated with carob molasses as a carbon source, with peptone as a nitrogen source; incubation was conducted at 37 °C for 96 h at an agitation speed of 300 rpm (114.95 mg/L). The HPLC analysis confirmed the synthesis of PHB by <i>B. paramycoides</i> to have a chromatogram retention time of 22.5 min. Carob waste was successfully valorized to PHB.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 3","pages":"318-337"},"PeriodicalIF":4.8,"publicationDate":"2022-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9576499/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40657937","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
AIMS MicrobiologyPub Date : 2022-07-15eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022022
Susan Nadin-Davis, Louise Pope, John Devenish, Ray Allain, Dele Ogunremi
{"title":"Evaluation of the use of CRISPR loci for discrimination of <i>Salmonella enterica</i> subsp. <i>enterica</i> serovar Enteritidis strains recovered in Canada and comparison with other subtyping methods.","authors":"Susan Nadin-Davis, Louise Pope, John Devenish, Ray Allain, Dele Ogunremi","doi":"10.3934/microbiol.2022022","DOIUrl":"10.3934/microbiol.2022022","url":null,"abstract":"<p><p><i>Salmonella enterica</i> subsp. <i>enterica</i> serovar Enteritidis remains one of the most important foodborne pathogens worldwide. To minimise its public health impact when outbreaks of the disease occur, timely investigation to identify and recall the contaminated food source is necessary. Central to this approach is the need for rapid and accurate identification of the bacterial subtype epidemiologically linked to the outbreak. While traditional methods of <i>S</i>. Enteritidis subtyping, such as pulsed field gel electrophoresis (PFGE) and phage typing (PT), have played an important role, the clonal nature of this organism has spurred efforts to improve subtyping resolution and timeliness through molecular based approaches. This study uses a cohort of 92 samples, recovered from a variety of sources, to compare these two traditional methods for <i>S</i>. Enteritidis subtyping with recently developed molecular techniques. These latter methods include the characterisation of two clustered regularly interspaced short palindromic repeats (CRISPR) loci, either in isolation or together with sequence analysis of virulence genes such as <i>fim</i>H. For comparison, another molecular technique developed in this laboratory involved the scoring of 60 informative single nucleotide polymorphisms (SNPs) distributed throughout the genome. Based on both the number of subtypes identified and Simpson's index of diversity, the CRISPR method was the least discriminatory and not significantly improved with the inclusion of <i>fim</i>H gene sequencing. While PT analysis identified the most subtypes, the SNP-PCR process generated the greatest index of diversity value. Combining methods consistently improved the number of subtypes identified, with the SNP/CRISPR typing scheme generating a level of diversity comparable with that of PT/PFGE. While these molecular methods, when combined, may have significant utility in real-world situations, this study suggests that CRISPR analysis alone lacks the discriminatory capability required to support investigations of foodborne disease outbreaks.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 3","pages":"300-317"},"PeriodicalIF":2.7,"publicationDate":"2022-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9576496/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40657933","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
AIMS MicrobiologyPub Date : 2022-07-12eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022021
Matthaios Papadimitriou-Olivgeris, Fevronia Kolonitsiou, Maria Militsopoulou, Iris Spiliopoulou, Nikolaos Giormezis
{"title":"Clonal dissemination and resistance genes among <i>Stenotrophomonas maltophilia</i> in a Greek University Hospital during a four-year period.","authors":"Matthaios Papadimitriou-Olivgeris, Fevronia Kolonitsiou, Maria Militsopoulou, Iris Spiliopoulou, Nikolaos Giormezis","doi":"10.3934/microbiol.2022021","DOIUrl":"https://doi.org/10.3934/microbiol.2022021","url":null,"abstract":"<p><p>Treatment of <i>Stenotrophomonas maltophilia</i> infections comprises of sulfamethoxazole/tripethoprim (SXT) or fluoroquinolones. We investigated antimicrobial resistance, presence of resistance genes (<i>sul1</i>, <i>smqnr</i>) and clonal dissemination in <i>S. maltophilia</i> from a university hospital. Among 62 isolates, 45 (73%) represented infection. Two isolates (3%) were resistant to SXT and three (5%) to levofloxacin. Twenty-nine isolates (47%), including two out of three levofloxacin-resistant, carried <i>smqnr</i>. Resistance of <i>S. maltophilia</i> was low and was not associated with <i>sul1</i> or <i>smqnr</i> carriage. Although high degree of genetic diversity was identified (29 pulsotypes), 22/62 (35.5%) strains were classified into four clones; clone b was associated with bacteraemias.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 3","pages":"292-299"},"PeriodicalIF":4.8,"publicationDate":"2022-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9576502/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40657939","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
AIMS MicrobiologyPub Date : 2022-07-08eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022020
Laura Weyersberg, Eva Klemens, Jule Buehler, Petra Vatter, Martin Hessling
{"title":"UVC, UVB and UVA susceptibility of Phi6 and its suitability as a SARS-CoV-2 surrogate.","authors":"Laura Weyersberg, Eva Klemens, Jule Buehler, Petra Vatter, Martin Hessling","doi":"10.3934/microbiol.2022020","DOIUrl":"10.3934/microbiol.2022020","url":null,"abstract":"<p><p>For SARS-CoV-2 disinfection systems or applications that are based on UVC, UVB or UVA irradiation, it would be desirable to have a SARS-CoV-2 surrogate for tests and development, which does not require a laboratory with a high biosafety level. The bacteriophage Phi 6, an enveloped RNA virus like coronaviruses, is an obvious candidate for such a surrogate. In this study, UVC, UVB and UVA log-reduction doses for Phi6 are determined by plaque assay. Log-reduction doses for SARS-CoV-2 are retrieved from a literature research. Because of a high variability of the published results, median log-reduction doses are determined for defined spectral ranges and compared to Phi6 data in the same intervals. The measured Phi6 log-reduction doses for UVC (254 nm), UVB (311 nm) and UVA (365 nm) are 31.7, 980 and 14 684 mJ/cm<sup>2</sup>, respectively. The determined median log-reduction doses for SARS-CoV-2 are much lower, only about 1.7 mJ/cm<sup>2</sup> within the spectral interval 251-270 nm. Therefore, Phi6 can be photoinactivated by all UV wavelengths but it is much less UV sensitive compared to SARS-CoV-2 in all UV spectral ranges. Thus, Phi6 is no convincing SARS-CoV-2 surrogate in UV applications.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 3","pages":"278-291"},"PeriodicalIF":2.7,"publicationDate":"2022-07-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9576498/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40657936","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
AIMS MicrobiologyPub Date : 2022-07-04eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022019
Ruba Mirghani, Tania Saba, Hebba Khaliq, Jennifer Mitchell, Lan Do, Liz Chambi, Kelly Diaz, Taylor Kennedy, Katia Alkassab, Thuhue Huynh, Mohamed Elmi, Jennifer Martinez, Suad Sawan, Girdhari Rijal
{"title":"Biofilms: Formation, drug resistance and alternatives to conventional approaches.","authors":"Ruba Mirghani, Tania Saba, Hebba Khaliq, Jennifer Mitchell, Lan Do, Liz Chambi, Kelly Diaz, Taylor Kennedy, Katia Alkassab, Thuhue Huynh, Mohamed Elmi, Jennifer Martinez, Suad Sawan, Girdhari Rijal","doi":"10.3934/microbiol.2022019","DOIUrl":"https://doi.org/10.3934/microbiol.2022019","url":null,"abstract":"<p><p>Biofilms are aggregates of bacteria, in most cases, which are resistant usually to broad-spectrum antibiotics in their typical concentrations or even in higher doses. A trend of increasing multi-drug resistance in biofilms, which are responsible for emerging life-threatening nosocomial infections, is becoming a serious problem. Biofilms, however, are at various sensitivity levels to environmental factors and are versatile in infectivity depending on virulence factors. This review presents the fundamental information about biofilms: formation, antibiotic resistance, impacts on public health and alternatives to conventional approaches. Novel developments in micro-biosystems that help reveal the new treatment tools by sensing and characterization of biofilms will also be discussed. Understanding the formation, structure, physiology and properties of biofilms better helps eliminate them by the usage of appropriate antibiotics or their control by novel therapy approaches, such as anti-biofilm molecules, effective gene editing, drug-delivery systems and probiotics.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 3","pages":"239-277"},"PeriodicalIF":4.8,"publicationDate":"2022-07-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9576500/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40657938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
AIMS MicrobiologyPub Date : 2022-06-24eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022018
Farhana Haque, Ishrat Jabeen, Chaman Ara Keya, Sabbir R Shuvo
{"title":"Whole-genome sequencing and comparative analysis of heavy metals tolerant <i>Bacillus anthracis</i> FHq strain isolated from tannery effluents in Bangladesh.","authors":"Farhana Haque, Ishrat Jabeen, Chaman Ara Keya, Sabbir R Shuvo","doi":"10.3934/microbiol.2022018","DOIUrl":"https://doi.org/10.3934/microbiol.2022018","url":null,"abstract":"<p><p>Heavy metal contamination of the environment is a primary concern in Bangladesh. This study aims to characterize a novel heavy metal tolerant strain, <i>Bacillus anthracis</i> FHq, isolated from the tannery effluents of Savar, Bangladesh. The strain could tolerate up to 5 mM of lead nitrate, 2.5 mM of sodium arsenate, chromium chloride, cobalt chloride, 1.5 mM cadmium acetate, and 1 mM of sodium arsenite. Whole-genome sequencing analysis revealed that the genome of the strain is around 5.2 Mbp long, and the G + C content is 35.4%. Besides, FHq has genes c<i>adC, zntA, arsCR, czcD</i>, and c<i>hrA</i>, which confer lead, arsenic, cobalt, and chromium resistance, respectively. A total of nineteen other closely related and completely sequenced <i>B. anthracis</i> strains were selected based on average nucleotide identity along with the FHq strain for phylogenomic and pan-genome analysis. The phylogenomic analysis predicted the inter-genomic evolutionary relationship of the strain isolated from Bangladesh, and it was closely related to a strain isolated from China. Pan-genome analysis revealed that the FHq strain possesses 6045 pan genes, 3802 core genes, and 152 unique genes in its genomic content. Hence, the genetic information and comparative analysis of the FHq strain might facilitate identifying the mechanisms conferring high resistance to lead in <i>B. anthracis</i> strains isolated from Bangladesh.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"227-239"},"PeriodicalIF":4.8,"publicationDate":"2022-06-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329874/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420432","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
AIMS MicrobiologyPub Date : 2022-06-17eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022017
Anastasia I Lavrova, Marine Z Dogonadze, Alexander V Sychev, Olga A Manicheva, Eugene B Postnikov
{"title":"Ensemble density-dependent synchronization of mycobacterial growth: BACTEC MGIT 960 fluorescence-based analysis and mathematical modelling of coupled biophysical and chemical processes.","authors":"Anastasia I Lavrova, Marine Z Dogonadze, Alexander V Sychev, Olga A Manicheva, Eugene B Postnikov","doi":"10.3934/microbiol.2022017","DOIUrl":"https://doi.org/10.3934/microbiol.2022017","url":null,"abstract":"<p><p>This study presents an analysis of <i>M. tuberculosis</i> growth data obtained using the BACTEC MGIT 960 system and respective mathematical models. The system is based on the detection of a decrease in oxygen level in the broth due to the bacterial respiration. It is shown that recordings sampled with a 1 hour rate provide an opportunity to distinguish between the oxygen consumption of growing cells and active cells division when the density of micro-organisms is sufficient to enter into the synchronized division mode. More specifically, the growth of culture is continuous only with large initial dilutions; otherwise, there are jumps between different growth stages with a time interval of 13-15 h. The combination of the oxygen-quenching kinetics for an analytic reagent and the population growth kinetics resulted in a mathematical model, which consists of mixing Verhulst's and Gompertz's models. The parameters of such mixing and switching between the models' prevalences are discussed with respect to oxygen uptake reactions reflected in the changes in the experimentally registered fluorescence level.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"208-226"},"PeriodicalIF":4.8,"publicationDate":"2022-06-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329876/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420431","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
AIMS MicrobiologyPub Date : 2022-05-16eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022016
Kholoud Baraka, Rania Abozahra, Maged Wasfy Helmy, Nada Salah El Dine El Meniawy, Sarah M Abdelhamid
{"title":"Investigation of the protective and therapeutic effects of <i>Lactobacillus casei</i> and <i>Saccharomyces cerevisiae</i> in a breast cancer mouse model.","authors":"Kholoud Baraka, Rania Abozahra, Maged Wasfy Helmy, Nada Salah El Dine El Meniawy, Sarah M Abdelhamid","doi":"10.3934/microbiol.2022016","DOIUrl":"10.3934/microbiol.2022016","url":null,"abstract":"<p><strong>Introduction: </strong>The development of novel strategies for cancer therapy is crucial to improve standard treatment protocols.</p><p><strong>Aim: </strong>This study aimed to determine the protective and therapeutic effects of heat-killed preparations of <i>Lactobacillus casei</i> and <i>Saccharomyces cerevisiae</i> in a breast cancer mouse model.</p><p><strong>Methods: </strong>Forty-two female BALB/c mice (7-8 weeks old) were divided into six groups (seven mice per group). Four groups were injected with 10<sup>7</sup> Ehrlich ascites tumor (EAT) cells suspended in phosphate-buffered saline (PBS) subcutaneously into the left side of the mammary fat pad. Tumor growth was monitored weekly until all animals developed a palpable tumor. The tumor-bearing mice in the experimental groups received heat-killed <i>L. casei</i> or <i>S. cerevisiae</i> three times per week for 35 days. The mice in the control group received PBS. The remaining two groups received heated <i>L. casei</i> or <i>S. cerevisiae</i> and then were injected with EAT cells. After 35 days, all mice were sacrificed to determine the immune response.</p><p><strong>Results: </strong>Animals that received heated <i>S. cerevisiae</i> exhibited the lowest rate of tumor growth compared with the other groups. TGF-β and IL-4 secretion was increased in all mice, whereas the secretion of INF-γ and IL-10 was decreased in breast tissues. Moreover, at the histopathological level, the volume of viable tumor in the control group was higher than in the treated groups.</p><p><strong>Conclusion: </strong>Supplementary treatment with <i>S. cerevisiae</i> resulted in the best outcome in the breast cancer model compared with other treated and vaccinated groups.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"193-207"},"PeriodicalIF":2.7,"publicationDate":"2022-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329878/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420436","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
AIMS MicrobiologyPub Date : 2022-05-09eCollection Date: 2022-01-01DOI: 10.3934/microbiol.2022015
Sanaz Dehbashi, Hamed Tahmasebi, Mohammad Yousef Alikhani, Fariba Keramat, Mohammad Reza Arabestani
{"title":"Optimization and development of high-resolution melting curve analysis (HRMA) assay for detection of New Delhi metallo-β-lactamase (NDM) producing <i>Pseudomonas aeruginosa</i>.","authors":"Sanaz Dehbashi, Hamed Tahmasebi, Mohammad Yousef Alikhani, Fariba Keramat, Mohammad Reza Arabestani","doi":"10.3934/microbiol.2022015","DOIUrl":"https://doi.org/10.3934/microbiol.2022015","url":null,"abstract":"<p><p>New Delhi metallo-β-lactamase-1 (NDM-1) producing <i>Pseudomonas aeruginosa</i> strain detection plays a vital role in confirming bacterial disease diagnosis and following the source of an outbreak for public health. However, the standard method for NDM-1 determination, which relies on the features of the colony of the bacteria cultured from the patient's specimen, is time-consuming and lacks accuracy and sensitivity. This study aimed to standardize a high-resolution melting curve analysis (HRMA) assay to detect NDM producing <i>P. aeruginosa</i>. For optimization and development of the HRMA method, a reference strain of <i>P. aeruginosa</i> was used. For evaluating the broad range PCR data, ABI Step One-Plus Manager Software version 3.2 and Precision Melt Analysis Software 3.02 (Applied Biosystems) were used. Based on the results, expected results were obtained for all tested strains, with high analytical sensitivity and specificity. Temperature melting analyses of the HRMA time PCR assays showed the Tm at 89.57 °C, 76.92 °C and 82.97 °C for N-1, N-2 and N-3 genes, respectively. Also, melting point temperatures of the <i>bla</i> <sub>VIM</sub>, <i>bla</i> <sub>SPM</sub> and <i>bla</i> <sub>SIM</sub> amplicons for isolates identified as MBL strains were 84.56 °C, 85.35 °C and 86.62 °C, respectively. The amplification results using negative control genomes as templates were negative, showing the specificity of the designed assays. Our study's data indicated that the sensitivity and specificity of the HRMA method are linked to the primer length and the fluorescent dye. We can further identify antibiotic resistance in NDMproducing <i>P. aeruginosa</i> by software analysis and melting curve analysis.</p>","PeriodicalId":46108,"journal":{"name":"AIMS Microbiology","volume":"8 2","pages":"178-192"},"PeriodicalIF":4.8,"publicationDate":"2022-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9329879/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"40420433","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}