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Matrix-Assisted Laser Desorption/Ionization Time Of Flight Mass Spectrometry (MALDI-TOF MS) technology is currently increasingly used in diagnostic laboratories as a cost effective, rapid and reliable routine technique for the identification and typing of microorganisms. In this study, we used MALDI-TOF MS to analyze a collection of 160 strains belonging to the Bacillus cereus group (57 B. anthracis, 49 B. cereus, 1 B. mycoides, 18 B. wiedmannii, 27 B. thuringiensis, 7 B. toyonensis and 1 B. weihenstephanensis) and to detect specific biomarkers which would allow an unequivocal identification. The Main Spectra Profiles (MSPs) were added to an in-house reference library, expanding the current commercial library which does not include B. toyonensis and B. wiedmannii mass spectra. The obtained mass spectra were statistically compared by Principal Component Analysis (PCA) that revealed seven different clusters. Moreover, for the identification purpose, were generated dedicate algorithms for a rapid and automatic detection of characteristic ion peaks after the mass spectra acquisition. The presence of specific biomarkers can be used to differentiate strains within the B. cereus group and to make a reliable identification of Bacillus anthracis, etiologic agent of anthrax, which is the most pathogenic and feared bacterium of the group. This could offer a critical time advantage for the diagnosis and for the clinical management of human anthrax even in case of bioterror attacks.
Viviana Manzulli; Valeria Rondinone; Alessandro Buchicchio; Luigina Serrecchia; Dora Cipolletta; Antonio Fasanella; Antonio Parisi; Laura Difato; Michela Iatarola; Angela Aceti; Elena Poppa; Francesco Tolve; Lorenzo Pace; Fiorenza Petruzzi; Ines Rovere; Donato Raele; Laura Del Sambro; Luigi Giangrossi; Domenico Galante. Discrimination of Bacillus cereus Group Members by MALDI-TOF Mass Spectrometry. Microorganisms 2021, 9, 1202 .
AMA StyleViviana Manzulli, Valeria Rondinone, Alessandro Buchicchio, Luigina Serrecchia, Dora Cipolletta, Antonio Fasanella, Antonio Parisi, Laura Difato, Michela Iatarola, Angela Aceti, Elena Poppa, Francesco Tolve, Lorenzo Pace, Fiorenza Petruzzi, Ines Rovere, Donato Raele, Laura Del Sambro, Luigi Giangrossi, Domenico Galante. Discrimination of Bacillus cereus Group Members by MALDI-TOF Mass Spectrometry. Microorganisms. 2021; 9 (6):1202.
Chicago/Turabian StyleViviana Manzulli; Valeria Rondinone; Alessandro Buchicchio; Luigina Serrecchia; Dora Cipolletta; Antonio Fasanella; Antonio Parisi; Laura Difato; Michela Iatarola; Angela Aceti; Elena Poppa; Francesco Tolve; Lorenzo Pace; Fiorenza Petruzzi; Ines Rovere; Donato Raele; Laura Del Sambro; Luigi Giangrossi; Domenico Galante. 2021. "Discrimination of Bacillus cereus Group Members by MALDI-TOF Mass Spectrometry." Microorganisms 9, no. 6: 1202.
In order to provide insights into the evolutionary and epidemiological viral dynamics during the current COVID-19 pandemic in South Eastern Italy, a total of 298 genomes of SARS-CoV-2 strains collected in the Apulia and Basilicata regions, between March 2020 and January 2021, were sequenced. The genomic analysis performed on the draft genomes allowed us to assign the genetic clades and lineages of belonging to each sample and provide an overview of the main circulating viral variants. Our data showed the spread in Apulia and Basilicata of SARS-CoV-2 variants which have emerged during the second wave of infections and are being currently monitored worldwide for their increased transmission rate and their possible impact on vaccines and therapies. These results emphasize the importance of genome sequencing for the epidemiological surveillance of the new SARS-CoV-2 variants’ spread.
Loredana Capozzi; Angelica Bianco; Laura Del Sambro; Domenico Simone; Antonio Lippolis; Maria Notarnicola; Graziano Pesole; Lorenzo Pace; Domenico Galante; Antonio Parisi. Genomic Surveillance of Circulating SARS-CoV-2 in South East Italy: A One-Year Retrospective Genetic Study. Viruses 2021, 13, 731 .
AMA StyleLoredana Capozzi, Angelica Bianco, Laura Del Sambro, Domenico Simone, Antonio Lippolis, Maria Notarnicola, Graziano Pesole, Lorenzo Pace, Domenico Galante, Antonio Parisi. Genomic Surveillance of Circulating SARS-CoV-2 in South East Italy: A One-Year Retrospective Genetic Study. Viruses. 2021; 13 (5):731.
Chicago/Turabian StyleLoredana Capozzi; Angelica Bianco; Laura Del Sambro; Domenico Simone; Antonio Lippolis; Maria Notarnicola; Graziano Pesole; Lorenzo Pace; Domenico Galante; Antonio Parisi. 2021. "Genomic Surveillance of Circulating SARS-CoV-2 in South East Italy: A One-Year Retrospective Genetic Study." Viruses 13, no. 5: 731.
In 2011, in Bangladesh, 11 anthrax outbreaks occurred in six districts of the country. Different types of samples were collected from May to September in the six districts where anthrax had occurred in order to detect and type Bacillus anthracis (B. anthracis) strains. Anthrax was detected in 46.6% of the samples analysed, in particular in soils, but also in bone samples, water, animal feed, and rumen ingesta of dead animals. Canonical single nucleotide polymorphisms (CanSNPs) analysis showed that all the isolates belonged to the major lineage A, sublineage A.Br.001/002 of China and Southeast Asia while the multi-locus variable number of tandem repeats (VNTRs) analysis (MLVA) with 15 VNTRs demonstrated the presence of five genotypes, of which two resulted to be new genotypes. The single nucleotide repeats (SNRs) analysis showed 13 SNR types; nevertheless, due to its higher discriminatory power, the presence of two isolates with different SNR-type polymorphisms was detected within two MLVA genotypes. This study assumes that soil is not the only reason for the spread of the disease in Bangladesh; contaminated feed and water can also play an important role in the epidemiology of anthrax. Possible explanations for these epidemiological relationships are discussed.
Domenico Galante; Viviana Manzulli; Luigina Serrecchia; Pietro Taranto; Martin Hugh-Jones; M. Hossain; Valeria Rondinone; Dora Cipolletta; Lorenzo Pace; Michela Iatarola; Francesco Tolve; Angela Aceti; Elena Poppa; Antonio Fasanella. Investigation on Anthrax in Bangladesh during the Outbreaks of 2011 and Definition of the Epidemiological Correlations. Pathogens 2021, 10, 481 .
AMA StyleDomenico Galante, Viviana Manzulli, Luigina Serrecchia, Pietro Taranto, Martin Hugh-Jones, M. Hossain, Valeria Rondinone, Dora Cipolletta, Lorenzo Pace, Michela Iatarola, Francesco Tolve, Angela Aceti, Elena Poppa, Antonio Fasanella. Investigation on Anthrax in Bangladesh during the Outbreaks of 2011 and Definition of the Epidemiological Correlations. Pathogens. 2021; 10 (4):481.
Chicago/Turabian StyleDomenico Galante; Viviana Manzulli; Luigina Serrecchia; Pietro Taranto; Martin Hugh-Jones; M. Hossain; Valeria Rondinone; Dora Cipolletta; Lorenzo Pace; Michela Iatarola; Francesco Tolve; Angela Aceti; Elena Poppa; Antonio Fasanella. 2021. "Investigation on Anthrax in Bangladesh during the Outbreaks of 2011 and Definition of the Epidemiological Correlations." Pathogens 10, no. 4: 481.
New SARS-CoV-2 mutations are constantly emerging, raising concerns of increased transmissibility, virulence or escape from host immune response. We describe a nested RT-PCR assay (~1500 bps) to detect multiple nucleotide changes resulting in key spike protein mutations distinctive of the major known circulating SARS-CoV-2 variants, including the three Variants of Concern (VOCs) 20I/501Y.V1 (United Kingdom), 20H/501Y.V2 (South Africa), and 20 J/501Y.V3 (Brazil), as well as the 20E.EU1 variant (Spain), the CAL.20C recently identified in California, and the mink-associated variant (GR, lineage B.1.1.298). Prior to application to field samples, the discriminatory potential of this PCR assay was explored using GISAID and Nextclade. To extend variant detection to challenging matrices such as sewage, where the amplification of long fragments is problematic, two short nested RT-PCR assays (~300 bps) were also designed, targeting portions of the region spanned by the long nested assay. The three newly-designed assays were then tested on field samples, including 31 clinical samples (7 fully-sequenced swab samples, and 24 uncharacterized ones) and 34 urban wastewater samples, some of which collected in areas where circulation of VOCs had been reported. The long assay successfully amplified 29 of the 31 swabs (93%), allowing the correct identification of variants 20I/501Y.V1 and 20E.EU1 present in the panel of previously characterized samples. The Spanish variant was detected in 14/24 of the uncharacterized samples as well. The sequences obtained using the short assays were consistent with those obtained with the long assay. Mutations characteristic of VOCs (UK and Brazilian variant) and of other variant (Spanish) were detected in sewage samples. To our knowledge, this is the first evidence of the presence of sequences harboring key mutations of 20I/501Y.V1 and 20 J/501Y.V3 in urban wastewaters, highlighting the potential contribution of wastewater surveillance to explore SARS-CoV-2 diversity. The developed nested RT-PCR assays can be used as an initial rapid screening test to select clinical samples containing mutations of interest. This can speed up diagnosis and optimize resources since it allows full genome sequencing to be done only on clinically relevant specimens. The assays can be also employed for a rapid and cost-effective detection of VOCs or other variants in sewage for the purposes of wastewater-based epidemiology. The approach proposed here can be used to better understand SARS-CoV-2 variant diversity, geographic distribution and impact worldwide.
G La Rosa; P. Mancini; G. Bonanno Ferraro; C. Veneri; M. Iaconelli; L. Lucentini; L. Bonadonna; S. Brusaferro; D. Brandtner; A. Fasanella; L. Pace; A. Parisi; D. Galante; E. Suffredini. Rapid screening for SARS-CoV-2 variants of concern in clinical and environmental samples using nested RT-PCR assays targeting key mutations of the spike protein. Water Research 2021, 197, 117104 -117104.
AMA StyleG La Rosa, P. Mancini, G. Bonanno Ferraro, C. Veneri, M. Iaconelli, L. Lucentini, L. Bonadonna, S. Brusaferro, D. Brandtner, A. Fasanella, L. Pace, A. Parisi, D. Galante, E. Suffredini. Rapid screening for SARS-CoV-2 variants of concern in clinical and environmental samples using nested RT-PCR assays targeting key mutations of the spike protein. Water Research. 2021; 197 ():117104-117104.
Chicago/Turabian StyleG La Rosa; P. Mancini; G. Bonanno Ferraro; C. Veneri; M. Iaconelli; L. Lucentini; L. Bonadonna; S. Brusaferro; D. Brandtner; A. Fasanella; L. Pace; A. Parisi; D. Galante; E. Suffredini. 2021. "Rapid screening for SARS-CoV-2 variants of concern in clinical and environmental samples using nested RT-PCR assays targeting key mutations of the spike protein." Water Research 197, no. : 117104-117104.
The coronavirus disease 2019 (Covid-19) pandemic is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and presents a global health emergency that needs urgent intervention. Viruses constantly change through mutation, and new variants of a virus are expected to occur over time. In the United Kingdom (UK), a new variant called B.1.1.7 has emerged with an unusually large number of mutations. The aim of this study is to evaluate the level of protection of sera from 12 patients infected and later healed in Apulia Region (Italy) with Covid-19 between March and November 2020, when the English variant was not circulating in this territory yet, against the new VOC 202012/01 variant by seroneutralization assay. The sera of patients had already been tested before, using a virus belonging to the lineage B.1 and showed an antibody neutralizing titer ranging between 1:160 and 1:320. All the 12 sera donors confirmed the same titers of neutralizing antibodies obtained with a strain belonging to the lineage B.1.1.7 (VOC 202012/01). These data indicate that antibodies produced in subjects infected with variants of Sars-CoV-2 strain before the appearance of the English one, seem to have a neutralizing power also against this variant.
Valeria Rondinone; Lorenzo Pace; Antonio Fasanella; Viviana Manzulli; Antonio Parisi; Maria Capobianchi; Angelo Ostuni; Maria Chironna; Elisabetta Caprioli; Maria Labonia; Dora Cipolletta; Ines Della Rovere; Luigina Serrecchia; Fiorenza Petruzzi; Germana Pennuzzi; Domenico Galante. VOC 202012/01 Variant Is Effectively Neutralized by Antibodies Produced by Patients Infected before Its Diffusion in Italy. Viruses 2021, 13, 276 .
AMA StyleValeria Rondinone, Lorenzo Pace, Antonio Fasanella, Viviana Manzulli, Antonio Parisi, Maria Capobianchi, Angelo Ostuni, Maria Chironna, Elisabetta Caprioli, Maria Labonia, Dora Cipolletta, Ines Della Rovere, Luigina Serrecchia, Fiorenza Petruzzi, Germana Pennuzzi, Domenico Galante. VOC 202012/01 Variant Is Effectively Neutralized by Antibodies Produced by Patients Infected before Its Diffusion in Italy. Viruses. 2021; 13 (2):276.
Chicago/Turabian StyleValeria Rondinone; Lorenzo Pace; Antonio Fasanella; Viviana Manzulli; Antonio Parisi; Maria Capobianchi; Angelo Ostuni; Maria Chironna; Elisabetta Caprioli; Maria Labonia; Dora Cipolletta; Ines Della Rovere; Luigina Serrecchia; Fiorenza Petruzzi; Germana Pennuzzi; Domenico Galante. 2021. "VOC 202012/01 Variant Is Effectively Neutralized by Antibodies Produced by Patients Infected before Its Diffusion in Italy." Viruses 13, no. 2: 276.
Background. The highly variable manifestation of the COVID-19 disease, from completely asymptomatic to fatal, is both a clinical and a public health issue. The criteria for discharge of hospitalized patients have been based so far on the negative result of Real-Time Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) tests, but the persistence of viral fragments may exceed that of the integral virus by weeks. The aim of our study was to verify the clearance of the virus at viral culture in patients hospitalized for COVID-19 that have clinically recovered but are still positive on nasopharyngeal swab. Methods. The study was conducted in hospitalized patients with positive RT-PCR on nasopharyngeal swab. Patients included were from asymptomatic to severe cases and performed nasopharyngeal control swabbing on day 14 for asymptomatic patient or at least three days after remission of symptoms. RT-PCR positive specimens were sent to a biosafety level 3 laboratory for viral culture. Results. We performed a combined analysis of RT-PCR and a highly sensitive in vitro culture from 84 samples of hospitalized patients. The average age was 46 ± 20.29, and 40.5% of the subjects had radiologically confirmed pneumonia, with average PaO2 of 72.35 ± 12.12and P/F ratio of 315 ± 83.15. Ct values for the N gene were lower in the first swab than in the control one (p < 0.001). The samples from 83 patients were negative at viral culture, and RT-PCR on the respective supernatants always confirmed the absence of viral growth. Conclusions. Our preliminary results demonstrate that patients clinically recovered for at least three days show the viral clearance at viral culture, and presumably they continued to not be contagious.
Viviana Manzulli; Giulia Scioscia; Giulio Giganti; Maria Rosaria Capobianchi; Donato Lacedonia; Lorenzo Pace; Dora Cipolletta; Pasquale Tondo; Rosella De Nittis; Valeria Rondinone; Luigina Serrecchia; Antonio Parisi; Domenico Galante; Sergio Lo Caputo; Teresa Antonia Santantonio; Damiana Moschetta; Vitangelo Dattoli; Antonio Fasanella; Maria Pia Foschino Barbaro. Real Time PCR and Culture-Based Virus Isolation Test in Clinically Recovered Patients: Is the Subject Still Infectious for SARS-CoV2? Journal of Clinical Medicine 2021, 10, 309 .
AMA StyleViviana Manzulli, Giulia Scioscia, Giulio Giganti, Maria Rosaria Capobianchi, Donato Lacedonia, Lorenzo Pace, Dora Cipolletta, Pasquale Tondo, Rosella De Nittis, Valeria Rondinone, Luigina Serrecchia, Antonio Parisi, Domenico Galante, Sergio Lo Caputo, Teresa Antonia Santantonio, Damiana Moschetta, Vitangelo Dattoli, Antonio Fasanella, Maria Pia Foschino Barbaro. Real Time PCR and Culture-Based Virus Isolation Test in Clinically Recovered Patients: Is the Subject Still Infectious for SARS-CoV2? Journal of Clinical Medicine. 2021; 10 (2):309.
Chicago/Turabian StyleViviana Manzulli; Giulia Scioscia; Giulio Giganti; Maria Rosaria Capobianchi; Donato Lacedonia; Lorenzo Pace; Dora Cipolletta; Pasquale Tondo; Rosella De Nittis; Valeria Rondinone; Luigina Serrecchia; Antonio Parisi; Domenico Galante; Sergio Lo Caputo; Teresa Antonia Santantonio; Damiana Moschetta; Vitangelo Dattoli; Antonio Fasanella; Maria Pia Foschino Barbaro. 2021. "Real Time PCR and Culture-Based Virus Isolation Test in Clinically Recovered Patients: Is the Subject Still Infectious for SARS-CoV2?" Journal of Clinical Medicine 10, no. 2: 309.
This report describes the evolution of COVID-19 in a 10 day-old-baby. The mother developed the disease immediately after childbirth and therefore a vertical transmission can be excluded. The isolation of the virus in cell culture with a cytopathic effect already visible after 48 h, indicates that the viral load of the newborn was quite high, but not serious course of the disease was observed. This paper wants to highlight the possible role of newborns and children in the spread of the disease.
Giuseppe Lenoci; Domenico Galante; Edmondo Ceci; Viviana Manzulli; Angela Maria Moramarco; Anna Chiaromonte; Giuseppina Labarile; Simone Lattarulo; Annalisa Resta; Lorenzo Pace; Valeria Rondinone; Antonio Parisi; Dora Cipolletta; Leonardo Marino; Iolanda Padalino; Luigina Serrecchia; Angela Aceti; Michela Iatarola; Francesco Tolve; Antonio Fasanella. Sars-CoV-2 isolation from a 10-day-old newborn in Italy: A case report. IDCases 2020, 22, e00960 -e00960.
AMA StyleGiuseppe Lenoci, Domenico Galante, Edmondo Ceci, Viviana Manzulli, Angela Maria Moramarco, Anna Chiaromonte, Giuseppina Labarile, Simone Lattarulo, Annalisa Resta, Lorenzo Pace, Valeria Rondinone, Antonio Parisi, Dora Cipolletta, Leonardo Marino, Iolanda Padalino, Luigina Serrecchia, Angela Aceti, Michela Iatarola, Francesco Tolve, Antonio Fasanella. Sars-CoV-2 isolation from a 10-day-old newborn in Italy: A case report. IDCases. 2020; 22 ():e00960-e00960.
Chicago/Turabian StyleGiuseppe Lenoci; Domenico Galante; Edmondo Ceci; Viviana Manzulli; Angela Maria Moramarco; Anna Chiaromonte; Giuseppina Labarile; Simone Lattarulo; Annalisa Resta; Lorenzo Pace; Valeria Rondinone; Antonio Parisi; Dora Cipolletta; Leonardo Marino; Iolanda Padalino; Luigina Serrecchia; Angela Aceti; Michela Iatarola; Francesco Tolve; Antonio Fasanella. 2020. "Sars-CoV-2 isolation from a 10-day-old newborn in Italy: A case report." IDCases 22, no. : e00960-e00960.