Validation and Implementation of a Diagnostic Algorithm for DNA Detection of Bordetella pertussis, B. parapertussis, and B. holmesii in a Pediatric Referral Hospital in Barcelona, Spain

Valero-Rello, Ana; Henares, Desirée; Acosta, Lesly; Jané, Mireia; Jordán, Iolanda; Godoy, Pere; Muñoz-Almagro, Carmen

doi:10.1128/jcm.01231-18

Cited by 21 publications

(14 citation statements)

References 27 publications

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“…It represents a crucial strategy for parasite load determination in clinical samples, and allows the number of parasites to be normalized by the DNA quantity present in the sample [15]. For human patients, the RNAse P is a commonly used target as an internal control of reaction [56][57][58][59][60][61][62]. In Leishmania however, there are no studies, except the present one, describing the use of an internal control to quantify parasite load from human patient's samples.…”

Section: Discussionmentioning

confidence: 99%

Comparison and clinical validation of qPCR assays targeting Leishmania 18S rDNA and HSP70 genes in patients with American Tegumentary Leishmaniasis

Filgueira¹,

Moreira²,

Cantanhêde³

et al. 2020

PLoS Negl Trop Dis

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Leishmaniasis is a worldwide neglected disease, encompassing asymptomatic infections and different clinical forms, such as American Tegumentary Leishmaniasis (ATL) which is part of the complex of diseases caused by protozoan parasites from Leishmania genus, transmitted by sand fly vectors. As a neglected disease, much effort is still needed in treatment and diagnosis. Currently, ATL diagnosis is mainly made by parasite detection by microscopy. The sensitivity of the method varies, and factors such as collection procedures interfere. Molecular approaches, specially based on Real Time PCR (qPCR) technique, has been widely used to detect Leishmania infection and to quantify parasite load, once it is a simple, rapid and sensitive methodology, capable to detect low parasite concentrations and less prone to variability. Although many studies have been already published addressing the use of this technique, an improvement on these methodologies, including an analytical validation, standardization and data association is demanded. Moreover, a proper validation by the assay by the use of clinical samples is still required. In this sense, the purpose of the present work is to compare the performance of qPCR using two commonly used targets (18S rDNA and HSP70) with an internal control (RNAse P) in multiplex reactions. Additionally, we validated reactions by assaying 88 samples from patients presenting different clinical forms of leishmaniasis (cutaneous, mucosal, recent and old lesions), representing the diversity found in Brazil's Amazon Region. Following the methodology proposed herein, the results indicate the use of both qPCR assays, 18S rDNA and HSP70, to achieve a very good net sensitivity (98.5%) and specificity (100%), performing simultaneous or sequential testing, respectively. With this approach, our main goal is to conclude the first step of a further multicenter study to propose the standardization of detection and quantification of Leishmania.

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Section: Discussionmentioning

confidence: 99%

Comparison and clinical validation of qPCR assays targeting Leishmania 18S rDNA and HSP70 genes in patients with American Tegumentary Leishmaniasis

Filgueira¹,

Moreira²,

Cantanhêde³

et al. 2020

PLoS Negl Trop Dis

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“…However, previous studies have found that B. holmesii infections are rare and mostly occur in older children and adults. Valero-Rello et al reported that B. holmesii was detected in only 0.9% (only in patients [ 4 years old) of patients suspected of pertussis [28]. Burgos-Rivera also reported a low positive rate of 0.66% [29].…”

Section: Discussionmentioning

confidence: 98%

Virus Coinfection is a Predictor of Radiologically Confirmed Pneumonia in Children with Bordetella pertussis Infection

Jiang

Chen

et al. 2020

Infect Dis Ther

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Introduction This study aimed to prospectively investigate the burden of pertussis in southeast Chinese children hospitalized with lower respiratory tract infection (LRTI) during a pertussis outbreak and to compare the outcomes of Bordetella pertussis infection with or without virus coinfections. Methods Children < 24 months of age hospitalized with LRTI were prospectively enrolled from January 2017 to December 2019. Demographic and clinical information were recorded, and respiratory tract samples were tested for the presence of B. pertussis and ten common viruses by polymerase chain reaction (PCR). Results Bordetella pertussis PCR was positive in 6.1% (202/4287) of the patients. Only 146 (72.3%) B. pertussis infections met the Centers for Disease Control and Prevention case definition for pertussis. Among the 202 subjects with B. pertussis infections, 81 (40.1%) were coinfected with at least 1 respiratory virus, with human rhinovirus being the most commonly detected virus (25.7%). No differences in clinical severity were observed between children with single B. pertussis infection and those with virus coinfection [odds ratio (OR) 0.75; 95% confidence interval (CI) 0.39–1.44]. However, children with virus coinfection were significantly more likely to present with radiologically confirmed pneumonia than those with a single B. pertussis infection (OR 2.62; CI 1.39–4.91). Conclusions Bordetella pertussis infection contributed to a high proportion of LRTI hospitalizations among southeast Chinese children. There were no significant differences in clinical severity between children with virus coinfection and single B. pertussis infection, although children coinfected with virus coinfection presented with pneumonia more frequently than those with single B. pertussis infection. Electronic supplementary material The online version of this article (10.1007/s40121-020-00376-5) contains supplementary material, which is available to authorized users.

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“…Current understanding of the whooping-cough etiology has changed, since it is not restricted to infections exclusively caused by B. pertussis , but embraces also pertussis-like illnesses caused by B. holmesii and B. parapertussis . B. pertussis is still a primary causative agent of whooping-cough, with B. holmesii , emerging as the second, and followed by B. parapertussis [ 10 , 12 , 13 , 28 ]. The relation between B. pertussis and B. holmesii infections remains unknown, but cases of the co-infection have been reported recently.…”

Section: Discussionmentioning

confidence: 99%

Bordetella holmesii Lipopolysaccharide Hide and Seek Game with Pertussis: Structural Analysis of the O-Specific Polysaccharide and the Core Oligosaccharide of the Type Strain ATCC 51541

Ucieklak

Koj

Niedziela

2020

IJMS

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Whooping cough is a highly contagious disease caused predominantly by Bordetella pertussis, but it also comprises of a pertussis-like illness caused by B. holmesii. The virulence factors of B. holmesii and their role in the pathogenesis remain unknown. Lipopolysaccharide is the main surface antigen of all Bordetellae. Data on the structural features of the lipopolysaccharide (LPS) of B. holmesii are scarce. The poly- and oligosaccharide components released by mild acidic hydrolysis of the LPS were separated and investigated by 1H and 13C NMR spectroscopy, mass spectrometry, and chemical methods. The structures of the O-specific polysaccharide and the core oligosaccharide of B. holmesii ATCC 51541 have been identified for the first time. The novel pentasaccharide repeating unit of the B. holmesii O-specific polysaccharide has the following structure: {→2)-α-l-Rhap-(1→6)-α-d-Glcp-(1→4)-[β-d-GlcpNAc-(1→3]-α-d-Galp-(1→3)-α-d-GlcpNAc-(1→}n. The SDS-PAGE and serological cross-reactivities of the B. holmesii LPS suggested the similarity between the core oligosaccharides of B. holmesii ATCC 51541 and B. pertussis strain 606. The main oligosaccharide fraction contained a nonasaccharide. The comparative analysis of the NMR spectra of B. holmesii core oligosaccharide fraction with this of the B. pertussis strain 606 indicated that the investigated core oligosaccharides were identical.

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Validation and Implementation of a Diagnostic Algorithm for DNA Detection of Bordetella pertussis, B. parapertussis, and B. holmesii in a Pediatric Referral Hospital in Barcelona, Spain

Cited by 21 publications

References 27 publications

Comparison and clinical validation of qPCR assays targeting Leishmania 18S rDNA and HSP70 genes in patients with American Tegumentary Leishmaniasis

Comparison and clinical validation of qPCR assays targeting Leishmania 18S rDNA and HSP70 genes in patients with American Tegumentary Leishmaniasis

Virus Coinfection is a Predictor of Radiologically Confirmed Pneumonia in Children with Bordetella pertussis Infection

Bordetella holmesii Lipopolysaccharide Hide and Seek Game with Pertussis: Structural Analysis of the O-Specific Polysaccharide and the Core Oligosaccharide of the Type Strain ATCC 51541

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