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Three years (2013–2016) of human respiratory syncytial virus surveillance at a tertiary hospital in Catalonia, Spain
Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
more, for monitoring of intensity, RSV-specific denominators are needed. http://dx.doi.org/10.1016/j.jcv.2016.08.234 Abstract no: 253 Presentation at ESCV 2016: Poster 195
Conclusions: Our results indicate that HSV 1-2 infection is frequent among patients hospitalized in intensive care unit. During the winter season this infection is linked to other respiratory viruses. The apparent clinical inefficiency of anti HSV 1-2 treatment indicates that the presence of the virus is more a witness of a clinically poor condition rather than a cause of it.
Herpes Simplex 1-2 in broncho alveolar fluid: A 5 years retrospective study
http://dx.doi.org/10.1016/j.jcv.2016.08.235
Catherine Mengelle 1,∗ , Jacques Izopet 1,2 , Emmanuel Bories 1 , Aurélie Lecour 1 , Jean-Michel Mansuy 1
Abstract no: 255 Presentation at ESCV 2016: Poster 196
1
Department of Virology, Toulouse University Hospital, Toulouse, France 2 Department of Physiopathology, Toulouse Purpan, Unité Inserm U563, Toulouse, France Aim: Respiratory viruses are very often detected in pneumonia thanks to uniplex and multiplex real-time PCR techniques. Moreover viruses that are not primilarly the cause of respiratory infections may also be detected such as HSV 1-2, varicella zoster virus and cytomegalovirus. The aim of our study was to analyze the prevalence of other respiratory infections among broncho-alveolar lavages (BAL) positive for HSV 1-2. The clinical outcome of patients according to the anti HSV 1-2 treatment was also analyzed. Material and methods: Data from hospitalized patients suffering from serious respiratory symptoms and whose bronchoalveolar lavages were positive for HSV 1-2 by an in-house real-time PCR were analyzed. Nucleic acids (NA) had been extracted with the MagNA Pure 96 DNA and Viral NA Small Volume kit® on the MagNA Pure 96TM instrument (Roche Molecular Diagnostics, Meylan, France). Samples had been tested for 16 respiratory viruses (influenza A and B, parainfluenza 1–4, respiratory syncytial viruses A and B, human metapneumovirus, coronaviruses 229E, OC43 and NL63, rhinoviruses, enteroviruses, bocaviruses and adenoviruses) with a multiplex RT-PCR (AnyplexTM II RV16 Detection® , Seegene) on the CFX96TM Real-Time System (Biorad diagnostics). NA had also been tested for cytomegalovirus, varicella zoster virus employing a monoplex in-house PCR on the Light Cycler or the Light Cycler 480TM (Roche Molecular Diagnostics, Meylan, France). Data were analyzed on StataTM software (StataCorp, Texas) using the exact Fisher test. Results: Between 2011 and 2015, 122 (73 males) patients attending an intensive care unit in the Toulouse University Hospital were selected with a HSV 1-2 positive result (mean age 62; 24–86). 119 samples were HSV-1 and three were HSV-2. 117 had been tested with the AnyplexTM II RV16 Detection Seegene® . All the viruses of the panel had been detected except parainfluenza 1, 2 and 4, human metapneumovirus and bocavirus; Influenza viruses were the most detected (n = 13), followed by rhinovirus (n = 6), respiratory syncytial virus (n = 5) and adenovirus (n = 4). 117 samples had been tested for cytomegalovirus (26 positive), and 90 for varicella zoster virus (negative). 28 among the 78 samples that tested positive for HSV 1-2 during the winter season (November to April) were also positive for another respiratory virus. During the summer season (May to October) 44 samples tested positive for HSV 1-2 with only 4 in coinfection (p < 0.005). The mortality rate did not differ between the HSV 1-2 positive patients treated with acyclovir or valacyclovir (n = 57) and those who were not (p = ns).
S117
Three years (2013–2016) of human respiratory syncytial virus surveillance at a tertiary hospital in Catalonia, Spain Laura Gimferrer ∗ , José Angel Rodrigo, Cristina Andrés, Isabel Dolores Oriolo, Maria Gema Codina, Paula Peremiquel, Maria del Carmen Martin, Francisco Fuentes, Rosario Saiz, Pilar Alcubilla, Magda Campins, Tomàs Pumarola, Andrés Antón Hospital Universitari Vall d’Hebron, Universitat Autònoma de Barcelona, Spain Background: Human respiratory syncytial virus (HRSV) is the most common respiratory pathogen and the main cause of lower respiratory tract infections among infants and young children. Its genome is a lineal single-stranded negative-sense RNA of approximately 15 kb that contains 10 genes encoding 11 proteins. The G glycoprotein in the viral envelope plays an essential role in the virus attachment. Antigenic and genetic differences in this protein lead classify HRSV into two different groups, HRSV-A and HRSV-B. Furthermore, based on the hypervariable region 2 (HVR-2) located in the C-terminal domain of the G protein several genotypes have been described. Selection pressure drives G protein to continuously evolve, resulting in the likely replacement of predominant genotype season by season. In the present study the epidemiology of HRSV viruses detected in respiratory specimens from patients attended at the Hospital Universitari Vall d’Hebron in Barcelona (Spain) during three consecutive years (from 2013 to 2016) has been described. Material and methods: From October 2013 (week 40) to March 2016 (week 20) respiratory specimens from patients were collected for laboratory confirmation of respiratory virus infection using immunochromatography (Binax Now RSV Card, Allere Scarborough Inc, USA), immunofluorescence (D3 Ultra 8TM DFA Respiratory Virus Screening & Identification Kit, Diagnostic HYBRIDS, USA) or real-time multiplex RT-PCR (Anyplex II RV16 Detection Kit, Seegene, Korea) assays. A nucleoprotein-specific real time RT-PCR was performed to determine HRSV group. In addition, phylogenetic analyses and molecular characterizations were carried out using MEGAv5.2 software based on the HVR-2 sequence from a representing sampling of HRSV per week. Results: A total of 16552 specimens were collected, of which 1324 (8.3%) were positive for HRSV. The virus showed a seasonal pattern of circulation, previous to influenza annual epidemics, with a maximum detection rates in the weeks 52 or 53 in all three seasons. Viruses belonging to both HRSV groups were detected: HRSV-A (662; 50%), HRSV-B (579, 44%), HRSV-A/B co-infection (8; <1%), and 75 (6%) remained unsubtyped. There was an alternation in the predominance of HRSV group by season; while HRSV-B was predominant during the first two seasons, HRSV-A became it during the third. Based on HVR-2 phylogenetic analyses, HRSV-A viruses belonged to ON1 genotype (153; 99%), but 2 (1%) to NA1.
S118
Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
In addition, HRSV-B viruses belonged to BA9 (153; 83%), BA10 (9; 5%) and an undefined BA (1; 1%). Nevertheless, 21 (11%) sequences from 2014 to 2015 season, closely related to the BA9 genotype, clustered together with a bootstrap value of 100% showing a pdistance between members of 0.006 and an average divergence within group of 0.004. Therefore, according to the criterion used by Venter et al. (J Gen Virol 2001; 82 (9): 2117-24), they might belong to a novel genotype (purposed name as BA13 in this study). However, viruses belonging to this new genotype were not found during the subsequent 2015–2016 season. Discussion and conclusions: Co-circulation of both HRSV groups has been reported during the three seasons. An alternation of the predominant HRSV group was shown during these three consecutive seasons. Although several genotypes were reported, the most of viruses belong to ON1 (HRSV-A) and BA9 (HRSV-B). The present study reports recent valuable data about the genetic diversity of circulating HRSV in the Southern Europe.
method, using primers specific for the amplification of the polymorphic sequence and two TaqMan-MGB probes specific for each allele (Express program and Genotyping assay service Applied Biosystem). The presence of at least one G allele (homo- or heterozygous) was significantly associated with overall disease severity (severity score 5–8), and related clinical parameters (but not with length of hospital stay, age or weight at hospital admission, weight at birth or gestational age, number of blood cells). Our previous study showed that infants carrying IL28B rs12979860 TT allele, that is in strong/moderate linkage disequilibrium with the IFNL4 G allele, had lower age at hospital admission, but did not suffer from a more severe bronchiolitis course. However, that study did not examine determinants of bronchiolitis severity in the RSV-infected children separately, because of a smaller number of samples. The present data suggest the importance of detecting IFNL4 SNPs in a larger group of infants affected with bronchiolitis. Further studies are needed also to understand the protective or detrimental effects of IFNL4 production during respiratory virus infections.
http://dx.doi.org/10.1016/j.jcv.2016.08.236 http://dx.doi.org/10.1016/j.jcv.2016.08.237 Abstract no: 263 Presentation at ESCV 2016: Poster 197 Evaluation of interferon lambda 4 nucleotide polymorphism in infants suffering from bronchiolitis A. Pierangeli 1,∗ , C. Scagnolari 1 , I. Calicchia 1 , I. Sciandra 1 , M. Gentile 1 , R. Nenna 2 , F. Midulla 2 , G. Antonelli 1 1
Department of Molecular Medicine, Sapienza University Rome, Italy 2 Department of Pediatrics, Sapienza University Rome, Italy The clinical spectrum of respiratory syncytial virus (RSV)associated bronchiolitis in infants is variable, ranging from a mild disease to a severe respiratory distress causing hospitalization. It is acquired that immune response and genetic heterogeneity of the host, together with well-known viral risk factors, contribute to RSV disease severity. Recent studies showed the importance of interferon (IFN) Lambda in the protection against RSV and other respiratory viruses. Our previous studies in bronchiolitis patients demonstrated higher mRNA levels of IFN Lambdas and of IFNstimulated genes in RSV-positive infants than in infants with HRV infection. Recently, it was shown that the novel ss469415590 SNP is more strongly associated with spontaneous HCV clearance and treatment-induced response than the IFN-lambda 3/IL28B SNP rs12979860. The ss469415590 SNP is a di-nucleotide mutant (TT > G) located in the region upstream IL28B gene; the unfavorable G allele is a frameshift variant creating the gene encoding a functional protein designated IFN-lambda 4 (IFNL4). Given the importance of the IFN lambda in respiratory infections, we sought to evaluate whether IFNL4 SNP could be associated with bronchiolitis severity. Hence, infants admitted to the Paediatric Department of Umberto I University Hospital, with a clinical diagnosis of bronchiolitis, were tested for ss469415590 SNP. Bronchiolitis severity was assessed with a score, based on respiratory rate, arterial oxygen saturation, presence of retractions and ability to feed (score range 0–8). For each sample, detection of 14 major respiratory viruses was performed and 122 samples positive only to RSV were selected for this preliminary study. DNA for the haplotype analysis was obtained from a buccal swab, when available or from archivial cell pellets from respiratory samples. TT/G genotyping was performed with the “StepOne Real-Time PCR System”
Abstract n◦ : 266 Presentation at ESCV 2016: Poster 198 Patterns of respiratory pathogen nasal colonization in the first year of life in healthy infants and infants with cystic fibrosis Insa Korten 1,∗ , Elisabeth Kieninger 1 , Njima Schläpfer 1 , Christine C. Ginocchio 2 , Carole Janis 3 , Shkipe Klenja 4 , Maria Teresa Barbani 4 , Urs Frey 5 , Nicolas Regamey 6 , Claudia Kuehni 7 , Markus Hilty 4 , Carmen Casaulta 1 , Philipp Latzin 1 , Meri Gorgievski 4 1
Division of Respiratory Medicine, Department of Pediatrics, Inselspital and University of Bern, Switzerland 2 Hofstra North Shore-LIJ School of Medicine, Hempstead, NY, USA 3 BioMérieuxSA, Verniolle, France 4 Institute for Infectious Diseases, University of Bern, Switzerland 5 University Children’s Hospital (UKBB), Basel, Switzerland 6 Division of Respiratory Medicine, Children’s Hospital Luzern, Switzerland 7 Institute for Social and Preventive Medicine, University of Bern, Switzerland Introduction and study aims: Respiratory infections are known to play a major role in morbidity and mortality, especially in early childhood and infancy. A number of studies have investigated pathogen colonization in otherwise healthy infants using PCR analysis of nasal swab material as an established diagnostic method. However, little is known about pathogen colonization in infants with chronic respiratory diseases like cystic fibrosis (CF). The aims of our study were: (1) to investigate feasibility and quality of parental collected nasal swab material for respiratory diagnostics; (2) to analyze possible differences in viral and atypical pathogen (Chlamydophila pnumoniae, Mycoplasma pneumoniae) colonization in healthy infants compared to infants with CF. Methods: 31 infants with CF and 32 unselected healthy infants were included in this prospective longitudinal study spanning the first year of life. Biweekly nasal FLOQSwabsTM (n = 1398) placed in UTM-RTTM (Copan, Italia) were collected by parents after
more, for monitoring of intensity, RSV-specific denominators are needed. http://dx.doi.org/10.1016/j.jcv.2016.08.234 Abstract no: 253 Presentation at ESCV 2016: Poster 195
Conclusions: Our results indicate that HSV 1-2 infection is frequent among patients hospitalized in intensive care unit. During the winter season this infection is linked to other respiratory viruses. The apparent clinical inefficiency of anti HSV 1-2 treatment indicates that the presence of the virus is more a witness of a clinically poor condition rather than a cause of it.
Herpes Simplex 1-2 in broncho alveolar fluid: A 5 years retrospective study
http://dx.doi.org/10.1016/j.jcv.2016.08.235
Catherine Mengelle 1,∗ , Jacques Izopet 1,2 , Emmanuel Bories 1 , Aurélie Lecour 1 , Jean-Michel Mansuy 1
Abstract no: 255 Presentation at ESCV 2016: Poster 196
1
Department of Virology, Toulouse University Hospital, Toulouse, France 2 Department of Physiopathology, Toulouse Purpan, Unité Inserm U563, Toulouse, France Aim: Respiratory viruses are very often detected in pneumonia thanks to uniplex and multiplex real-time PCR techniques. Moreover viruses that are not primilarly the cause of respiratory infections may also be detected such as HSV 1-2, varicella zoster virus and cytomegalovirus. The aim of our study was to analyze the prevalence of other respiratory infections among broncho-alveolar lavages (BAL) positive for HSV 1-2. The clinical outcome of patients according to the anti HSV 1-2 treatment was also analyzed. Material and methods: Data from hospitalized patients suffering from serious respiratory symptoms and whose bronchoalveolar lavages were positive for HSV 1-2 by an in-house real-time PCR were analyzed. Nucleic acids (NA) had been extracted with the MagNA Pure 96 DNA and Viral NA Small Volume kit® on the MagNA Pure 96TM instrument (Roche Molecular Diagnostics, Meylan, France). Samples had been tested for 16 respiratory viruses (influenza A and B, parainfluenza 1–4, respiratory syncytial viruses A and B, human metapneumovirus, coronaviruses 229E, OC43 and NL63, rhinoviruses, enteroviruses, bocaviruses and adenoviruses) with a multiplex RT-PCR (AnyplexTM II RV16 Detection® , Seegene) on the CFX96TM Real-Time System (Biorad diagnostics). NA had also been tested for cytomegalovirus, varicella zoster virus employing a monoplex in-house PCR on the Light Cycler or the Light Cycler 480TM (Roche Molecular Diagnostics, Meylan, France). Data were analyzed on StataTM software (StataCorp, Texas) using the exact Fisher test. Results: Between 2011 and 2015, 122 (73 males) patients attending an intensive care unit in the Toulouse University Hospital were selected with a HSV 1-2 positive result (mean age 62; 24–86). 119 samples were HSV-1 and three were HSV-2. 117 had been tested with the AnyplexTM II RV16 Detection Seegene® . All the viruses of the panel had been detected except parainfluenza 1, 2 and 4, human metapneumovirus and bocavirus; Influenza viruses were the most detected (n = 13), followed by rhinovirus (n = 6), respiratory syncytial virus (n = 5) and adenovirus (n = 4). 117 samples had been tested for cytomegalovirus (26 positive), and 90 for varicella zoster virus (negative). 28 among the 78 samples that tested positive for HSV 1-2 during the winter season (November to April) were also positive for another respiratory virus. During the summer season (May to October) 44 samples tested positive for HSV 1-2 with only 4 in coinfection (p < 0.005). The mortality rate did not differ between the HSV 1-2 positive patients treated with acyclovir or valacyclovir (n = 57) and those who were not (p = ns).
S117
Three years (2013–2016) of human respiratory syncytial virus surveillance at a tertiary hospital in Catalonia, Spain Laura Gimferrer ∗ , José Angel Rodrigo, Cristina Andrés, Isabel Dolores Oriolo, Maria Gema Codina, Paula Peremiquel, Maria del Carmen Martin, Francisco Fuentes, Rosario Saiz, Pilar Alcubilla, Magda Campins, Tomàs Pumarola, Andrés Antón Hospital Universitari Vall d’Hebron, Universitat Autònoma de Barcelona, Spain Background: Human respiratory syncytial virus (HRSV) is the most common respiratory pathogen and the main cause of lower respiratory tract infections among infants and young children. Its genome is a lineal single-stranded negative-sense RNA of approximately 15 kb that contains 10 genes encoding 11 proteins. The G glycoprotein in the viral envelope plays an essential role in the virus attachment. Antigenic and genetic differences in this protein lead classify HRSV into two different groups, HRSV-A and HRSV-B. Furthermore, based on the hypervariable region 2 (HVR-2) located in the C-terminal domain of the G protein several genotypes have been described. Selection pressure drives G protein to continuously evolve, resulting in the likely replacement of predominant genotype season by season. In the present study the epidemiology of HRSV viruses detected in respiratory specimens from patients attended at the Hospital Universitari Vall d’Hebron in Barcelona (Spain) during three consecutive years (from 2013 to 2016) has been described. Material and methods: From October 2013 (week 40) to March 2016 (week 20) respiratory specimens from patients were collected for laboratory confirmation of respiratory virus infection using immunochromatography (Binax Now RSV Card, Allere Scarborough Inc, USA), immunofluorescence (D3 Ultra 8TM DFA Respiratory Virus Screening & Identification Kit, Diagnostic HYBRIDS, USA) or real-time multiplex RT-PCR (Anyplex II RV16 Detection Kit, Seegene, Korea) assays. A nucleoprotein-specific real time RT-PCR was performed to determine HRSV group. In addition, phylogenetic analyses and molecular characterizations were carried out using MEGAv5.2 software based on the HVR-2 sequence from a representing sampling of HRSV per week. Results: A total of 16552 specimens were collected, of which 1324 (8.3%) were positive for HRSV. The virus showed a seasonal pattern of circulation, previous to influenza annual epidemics, with a maximum detection rates in the weeks 52 or 53 in all three seasons. Viruses belonging to both HRSV groups were detected: HRSV-A (662; 50%), HRSV-B (579, 44%), HRSV-A/B co-infection (8; <1%), and 75 (6%) remained unsubtyped. There was an alternation in the predominance of HRSV group by season; while HRSV-B was predominant during the first two seasons, HRSV-A became it during the third. Based on HVR-2 phylogenetic analyses, HRSV-A viruses belonged to ON1 genotype (153; 99%), but 2 (1%) to NA1.
S118
Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
In addition, HRSV-B viruses belonged to BA9 (153; 83%), BA10 (9; 5%) and an undefined BA (1; 1%). Nevertheless, 21 (11%) sequences from 2014 to 2015 season, closely related to the BA9 genotype, clustered together with a bootstrap value of 100% showing a pdistance between members of 0.006 and an average divergence within group of 0.004. Therefore, according to the criterion used by Venter et al. (J Gen Virol 2001; 82 (9): 2117-24), they might belong to a novel genotype (purposed name as BA13 in this study). However, viruses belonging to this new genotype were not found during the subsequent 2015–2016 season. Discussion and conclusions: Co-circulation of both HRSV groups has been reported during the three seasons. An alternation of the predominant HRSV group was shown during these three consecutive seasons. Although several genotypes were reported, the most of viruses belong to ON1 (HRSV-A) and BA9 (HRSV-B). The present study reports recent valuable data about the genetic diversity of circulating HRSV in the Southern Europe.
method, using primers specific for the amplification of the polymorphic sequence and two TaqMan-MGB probes specific for each allele (Express program and Genotyping assay service Applied Biosystem). The presence of at least one G allele (homo- or heterozygous) was significantly associated with overall disease severity (severity score 5–8), and related clinical parameters (but not with length of hospital stay, age or weight at hospital admission, weight at birth or gestational age, number of blood cells). Our previous study showed that infants carrying IL28B rs12979860 TT allele, that is in strong/moderate linkage disequilibrium with the IFNL4 G allele, had lower age at hospital admission, but did not suffer from a more severe bronchiolitis course. However, that study did not examine determinants of bronchiolitis severity in the RSV-infected children separately, because of a smaller number of samples. The present data suggest the importance of detecting IFNL4 SNPs in a larger group of infants affected with bronchiolitis. Further studies are needed also to understand the protective or detrimental effects of IFNL4 production during respiratory virus infections.
http://dx.doi.org/10.1016/j.jcv.2016.08.236 http://dx.doi.org/10.1016/j.jcv.2016.08.237 Abstract no: 263 Presentation at ESCV 2016: Poster 197 Evaluation of interferon lambda 4 nucleotide polymorphism in infants suffering from bronchiolitis A. Pierangeli 1,∗ , C. Scagnolari 1 , I. Calicchia 1 , I. Sciandra 1 , M. Gentile 1 , R. Nenna 2 , F. Midulla 2 , G. Antonelli 1 1
Department of Molecular Medicine, Sapienza University Rome, Italy 2 Department of Pediatrics, Sapienza University Rome, Italy The clinical spectrum of respiratory syncytial virus (RSV)associated bronchiolitis in infants is variable, ranging from a mild disease to a severe respiratory distress causing hospitalization. It is acquired that immune response and genetic heterogeneity of the host, together with well-known viral risk factors, contribute to RSV disease severity. Recent studies showed the importance of interferon (IFN) Lambda in the protection against RSV and other respiratory viruses. Our previous studies in bronchiolitis patients demonstrated higher mRNA levels of IFN Lambdas and of IFNstimulated genes in RSV-positive infants than in infants with HRV infection. Recently, it was shown that the novel ss469415590 SNP is more strongly associated with spontaneous HCV clearance and treatment-induced response than the IFN-lambda 3/IL28B SNP rs12979860. The ss469415590 SNP is a di-nucleotide mutant (TT > G) located in the region upstream IL28B gene; the unfavorable G allele is a frameshift variant creating the gene encoding a functional protein designated IFN-lambda 4 (IFNL4). Given the importance of the IFN lambda in respiratory infections, we sought to evaluate whether IFNL4 SNP could be associated with bronchiolitis severity. Hence, infants admitted to the Paediatric Department of Umberto I University Hospital, with a clinical diagnosis of bronchiolitis, were tested for ss469415590 SNP. Bronchiolitis severity was assessed with a score, based on respiratory rate, arterial oxygen saturation, presence of retractions and ability to feed (score range 0–8). For each sample, detection of 14 major respiratory viruses was performed and 122 samples positive only to RSV were selected for this preliminary study. DNA for the haplotype analysis was obtained from a buccal swab, when available or from archivial cell pellets from respiratory samples. TT/G genotyping was performed with the “StepOne Real-Time PCR System”
Abstract n◦ : 266 Presentation at ESCV 2016: Poster 198 Patterns of respiratory pathogen nasal colonization in the first year of life in healthy infants and infants with cystic fibrosis Insa Korten 1,∗ , Elisabeth Kieninger 1 , Njima Schläpfer 1 , Christine C. Ginocchio 2 , Carole Janis 3 , Shkipe Klenja 4 , Maria Teresa Barbani 4 , Urs Frey 5 , Nicolas Regamey 6 , Claudia Kuehni 7 , Markus Hilty 4 , Carmen Casaulta 1 , Philipp Latzin 1 , Meri Gorgievski 4 1
Division of Respiratory Medicine, Department of Pediatrics, Inselspital and University of Bern, Switzerland 2 Hofstra North Shore-LIJ School of Medicine, Hempstead, NY, USA 3 BioMérieuxSA, Verniolle, France 4 Institute for Infectious Diseases, University of Bern, Switzerland 5 University Children’s Hospital (UKBB), Basel, Switzerland 6 Division of Respiratory Medicine, Children’s Hospital Luzern, Switzerland 7 Institute for Social and Preventive Medicine, University of Bern, Switzerland Introduction and study aims: Respiratory infections are known to play a major role in morbidity and mortality, especially in early childhood and infancy. A number of studies have investigated pathogen colonization in otherwise healthy infants using PCR analysis of nasal swab material as an established diagnostic method. However, little is known about pathogen colonization in infants with chronic respiratory diseases like cystic fibrosis (CF). The aims of our study were: (1) to investigate feasibility and quality of parental collected nasal swab material for respiratory diagnostics; (2) to analyze possible differences in viral and atypical pathogen (Chlamydophila pnumoniae, Mycoplasma pneumoniae) colonization in healthy infants compared to infants with CF. Methods: 31 infants with CF and 32 unselected healthy infants were included in this prospective longitudinal study spanning the first year of life. Biweekly nasal FLOQSwabsTM (n = 1398) placed in UTM-RTTM (Copan, Italia) were collected by parents after