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1.
Arch Virol ; 166(6): 1599-1605, 2021 Jun.
Article in English | MEDLINE | ID: covidwho-1549420

ABSTRACT

Pigeon paramyxovirus-1 (PPMV-1) is a strain of Newcastle disease virus (NDV) that has adapted to infect pigeons and poses a constant threat to the commercial poultry industry. Early detection via rapid and sensitive methods, along with timely preventative and mitigating actions, is important for reducing the spread of PPMV-1. Here, we report the development of a TaqMan loop-mediated isothermal amplification assay (TaqMan-LAMP) for rapid and specific detection of PPMV-1 based on the F gene. This system makes use of six novel primers and a TaqMan probe that targets nine distinct regions of the F gene that are highly conserved among PPMV-1 isolates. The results showed that the limit of detection was 10 copies µL-1 for PPMV-1 cDNA and 0.1 ng for PPMV-1 RNA. The reaction was completed within 25 min and was thus faster than conventional RT-PCR. Moreover, no cross-reactions with similar viruses or with peste des petits ruminants virus (PPRV) or NDV LaSota vaccine strains were observed under the same conditions. To evaluate the applicability of the assay, the TaqMan-LAMP assay and a commercial RT-PCR assay were compared using 108 clinical samples, and the concordance rate between two methods was found to be 96.3%. The newly developed PPMV-1 TaqMan-LAMP assay can therefore be used for simple, efficient, rapid, specific, and sensitive diagnosis of PPMV-1 infections.


Subject(s)
Molecular Diagnostic Techniques/veterinary , Newcastle disease virus/genetics , Newcastle disease virus/isolation & purification , Nucleic Acid Amplification Techniques/veterinary , Animals , Columbidae , Feces/virology , Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , RNA, Viral , Sensitivity and Specificity , Time Factors
3.
Eur Rev Med Pharmacol Sci ; 25(21): 6745-6766, 2021 11.
Article in English | MEDLINE | ID: covidwho-1524863

ABSTRACT

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that has created the current pandemic, has caused a worldwide worry. Different countries have since enforced varying levels of lockdowns and guidelines for their populations to follow in a serious effort to mitigate the spread. Up until recently, the majority of these regulations and policies were established on the assumption that the dominant routes of transmission of this virus are through droplets and fomite contact. However, there is now a substantial amount of research pointing towards the strong possibility that SARS-CoV-2 can spread through airborne means. The World Health Organization (WHO) and the Center for Disease Control and Prevention (CDC) have recently recognized this, which poses the question of whether our collective methods of lessening transmission risk and keeping people safe have been sufficient. This paper is a comprehensive review of the evidence on SARS-CoV-2 being an airborne disease, through different epidemiological, experimental, and animal-model based published research. Studies opposing this evidence have also been discussed. The majority of these studies are favoring the high plausibility of SARS-CoV-2 aerosol transmission, and therefore the many implications of aerosol transmission have been discussed in this paper to suggest effective mitigation and control strategies.


Subject(s)
Aerosols , COVID-19/transmission , SARS-CoV-2/physiology , Animals , COVID-19/epidemiology , COVID-19/prevention & control , COVID-19/virology , Cricetinae , Disease Models, Animal , Equipment and Supplies, Hospital/virology , Feces/virology , Humans , Masks , Pandemics , Particulate Matter , RNA, Viral/metabolism , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Ventilation/standards , Virus Inactivation
4.
J Korean Med Sci ; 36(44): e301, 2021 Nov 15.
Article in English | MEDLINE | ID: covidwho-1526760

ABSTRACT

We used serial rectal swabs to investigate the amount and duration of virus secretion through the gastrointestinal tract and assessed the association between fecal shedding and gastrointestinal symptoms and to clarify the clinical usefulness testing rectal swabs. We enrolled ten adult patients hospitalized with symptomatic coronavirus disease 2019 (COVID-19). Respiratory and stool specimens were collected by physicians. The presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was confirmed using real-time reverse-transcription polymerase chain reaction. All ten patients had respiratory symptoms, six had diarrhea, and seven were positive for SARS-CoV-2 on rectal swabs. The viral loads in the respiratory specimens was higher than those in the rectal specimens, and no rectal specimens were positive after the respiratory specimens became negative. There was no association between gastrointestinal symptoms, pneumonia, severity, and rectal viral load. Rectal swabs may play a role in detecting SARS-CoV-2 in individuals with suspected COVID-19, regardless of gastrointestinal symptoms.


Subject(s)
COVID-19 Nucleic Acid Testing/methods , COVID-19/virology , Rectum/virology , SARS-CoV-2/isolation & purification , Virus Shedding , Adolescent , Adult , Aged , Aged, 80 and over , COVID-19/complications , COVID-19/transmission , Diarrhea/etiology , Diarrhea/virology , Feces/virology , Female , Humans , Male , Middle Aged , Nasopharynx/virology , Prospective Studies , Real-Time Polymerase Chain Reaction , Severity of Illness Index , Time Factors , Viral Load
5.
Viruses ; 12(8)2020 08 18.
Article in English | MEDLINE | ID: covidwho-1453290

ABSTRACT

Enteric viral co-infections, infections involving more than one virus, have been reported for a diverse group of etiological agents, including rotavirus, norovirus, astrovirus, adenovirus, and enteroviruses. These pathogens are causative agents for acute gastroenteritis and diarrheal disease in immunocompetent and immunocompromised individuals of all ages globally. Despite virus-virus co-infection events in the intestine being increasingly detected, little is known about their impact on disease outcomes or human health. Here, we review what is currently known about the clinical prevalence of virus-virus co-infections and how co-infections may influence vaccine responses. While experimental investigations into enteric virus co-infections have been limited, we highlight in vivo and in vitro models with exciting potential to investigate viral co-infections. Many features of virus-virus co-infection mechanisms in the intestine remain unclear, and further research will be critical.


Subject(s)
Coinfection/virology , Gastroenteritis/virology , Virus Diseases/physiopathology , Viruses/classification , Viruses/pathogenicity , Animals , Asymptomatic Infections , Disease Models, Animal , Feces/virology , Humans , Intestines/virology , Mice , Primates
6.
Viruses ; 13(10)2021 10 06.
Article in English | MEDLINE | ID: covidwho-1460085

ABSTRACT

According to various estimates, only a small percentage of existing viruses have been discovered, naturally much less being represented in the genomic databases. High-throughput sequencing technologies develop rapidly, empowering large-scale screening of various biological samples for the presence of pathogen-associated nucleotide sequences, but many organisms are yet to be attributed specific loci for identification. This problem particularly impedes viral screening, due to vast heterogeneity in viral genomes. In this paper, we present a new bioinformatic pipeline, VirIdAl, for detecting and identifying viral pathogens in sequencing data. We also demonstrate the utility of the new software by applying it to viral screening of the feces of bats collected in the Moscow region, which revealed a significant variety of viruses associated with bats, insects, plants, and protozoa. The presence of alpha and beta coronavirus reads, including the MERS-like bat virus, deserves a special mention, as it once again indicates that bats are indeed reservoirs for many viral pathogens. In addition, it was shown that alignment-based methods were unable to identify the taxon for a large proportion of reads, and we additionally applied other approaches, showing that they can further reveal the presence of viral agents in sequencing data. However, the incompleteness of viral databases remains a significant problem in the studies of viral diversity, and therefore necessitates the use of combined approaches, including those based on machine learning methods.


Subject(s)
Alphacoronavirus/isolation & purification , Betacoronavirus/isolation & purification , Chiroptera/virology , Genome, Viral/genetics , Metagenome/genetics , Alphacoronavirus/classification , Alphacoronavirus/genetics , Animals , Betacoronavirus/classification , Betacoronavirus/genetics , Chiroptera/genetics , Computational Biology/methods , Feces/virology , High-Throughput Nucleotide Sequencing , Metagenomics/methods , Moscow , Phycodnaviridae/classification , Phycodnaviridae/genetics , Phycodnaviridae/isolation & purification , Sequence Analysis, DNA
8.
Nat Commun ; 12(1): 5753, 2021 10 01.
Article in English | MEDLINE | ID: covidwho-1447302

ABSTRACT

Patients with COVID-19 shed SARS-CoV-2 RNA in stool, sometimes well after their respiratory infection has cleared. This may be significant for patient health, epidemiology, and diagnosis. However, methods to preserve stool, and to extract and quantify viral RNA are not standardized. We test the performance of three preservative approaches at yielding detectable SARS-CoV-2 RNA: the OMNIgene-GUT kit, Zymo DNA/RNA shield kit, and the most commonly applied, storage without preservative. We test these in combination with three extraction kits: QIAamp Viral RNA Mini Kit, Zymo Quick-RNA Viral Kit, and MagMAX Viral/Pathogen Kit. We also test the utility of ddPCR and RT-qPCR for the reliable quantification of SARS-CoV-2 RNA from stool. We identify that the Zymo DNA/RNA preservative and the QiaAMP extraction kit yield more detectable RNA than the others, using both ddPCR and RT-qPCR. Taken together, we recommend a comprehensive methodology for preservation, extraction and detection of RNA from SARS-CoV-2 and other coronaviruses in stool.


Subject(s)
COVID-19 Nucleic Acid Testing/standards , Feces/virology , SARS-CoV-2/isolation & purification , COVID-19/diagnosis , Coronavirus Nucleocapsid Proteins/genetics , Humans , Phosphoproteins/genetics , Preservation, Biological/standards , RNA, Viral/analysis , RNA, Viral/genetics , Reagent Kits, Diagnostic , Reference Standards , SARS-CoV-2/genetics , Specimen Handling/standards , Viral Load/standards
9.
Pol J Microbiol ; 70(3): 401-404, 2021 Sep.
Article in English | MEDLINE | ID: covidwho-1441450

ABSTRACT

SARS-CoV-2 was found in a recovered patient's stool specimen by combining quantitative reverse transcription PCR (qRT-PCR) and genome sequencing. The patient was virus positive in stool specimens for at least an additional 15 days after he was recovered, whereas respiratory tract specimens were negative. The discovery of the complete genome of SARS-CoV-2 in the stool sample of the recovered patient demonstrates a cautionary warning that the potential mode of the virus transmission cannot be excluded through the fecal-oral route after viral clearance in the respiratory tract.


Subject(s)
COVID-19/virology , Convalescence , Feces/virology , Genome, Viral , SARS-CoV-2/genetics , Whole Genome Sequencing , Adult , COVID-19/diagnostic imaging , COVID-19/transmission , China , Cough/virology , Fever/virology , Humans , Male , SARS-CoV-2/isolation & purification , Tomography, X-Ray Computed
10.
J Pediatr ; 237: 136-142, 2021 Oct.
Article in English | MEDLINE | ID: covidwho-1426917

ABSTRACT

OBJECTIVE: To evaluate the role of childcare facilities in the transmission of severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) in a longitudinal study to gain further knowledge of SARS-CoV-2 prevalence, transmission, and spread among preschool children, their parents, and their caregivers. STUDY DESIGN: Children aged 1-6 years, their parents, and their caregivers in 14 childcare facilities in Dresden, Saxony/Germany were invited to participate in the KiTaCoviDD19-study between July 2020 and January 2021. Seroprevalence of SARS-CoV-2 antibodies was assessed up to 4 times during the study period in all participating adults, and demographic characteristics, as well as epidemiologic information on personal SARS-CoV-2 history were obtained. Samples for stool virus shedding of SARS-CoV-2 were analyzed by polymerase chain reaction every 2-4 weeks in all participating children. RESULTS: In total, 318 children, 299 parents and 233 childcare workers were enrolled. By January 2021, 11% of the participating adults were found to be seropositive, whereas the percentage of children shedding SARS-CoV-2 was 6.8%. Overall, we detected 17 children with SARS-CoV-2 virus shedding in 8 different childcare facilities. In 4 facilities, there were a maximum of 3 connected cases in children. Approximately 50% of SARS-CoV-2 infections in the children could not be connected to a secondary case in our study population. CONCLUSIONS: This study does not provide evidence of relevant asymptomatic ("silent") spread of SARS-CoV-2 in childcare facilities in both low- and high-prevalence settings. Our findings add to the evidence that childcare and educational settings do not have a crucial role in driving the SARS-CoV-2 pandemic.


Subject(s)
COVID-19/transmission , Child Day Care Centers/statistics & numerical data , Adult , COVID-19/epidemiology , COVID-19 Testing/methods , Child , Child, Preschool , Feces/virology , Female , Germany/epidemiology , Humans , Infant , Longitudinal Studies , Male , Middle Aged , Pandemics , Parents , Prevalence , Quarantine , Retrospective Studies , SARS-CoV-2 , Virus Shedding
13.
BMC Microbiol ; 21(1): 242, 2021 09 06.
Article in English | MEDLINE | ID: covidwho-1398839

ABSTRACT

BACKGROUND: SARS-CoV-2 has been detected in stool samples of COVID-19 patients, with potential implications for faecal-oral transmission. Compared to nasopharyngeal swab samples, the complexity of the stool matrix poses a challenge in the detection of the virus that has not yet been solved. However, robust and reliable methods are needed to estimate the prevalence and persistence of SARS-CoV-2 in the gut and to ensure the safety of microbiome-based procedures such as faecal microbiota transplant (FMT). The aim of this study was to establish a sensitive and reliable method for detecting SARS-CoV-2 in stool samples. RESULTS: Stool samples from individuals free of SARS-CoV-2 were homogenised in saline buffer and spiked with a known titre of inactivated virus ranging from 50 to 750 viral particles per 100 mg stool. Viral particles were concentrated by ultrafiltration, RNA was extracted, and SARS-CoV-2 was detected via real-time reverse-transcription polymerase chain reaction (RT-qPCR) using the CDC primers and probes. The RNA extraction procedure we used allowed for the detection of SARS-CoV-2 via RT-qPCR in most of the stool samples tested. We could detect as few as 50 viral particles per 100 mg of stool. However, high variability was observed across samples at low viral titres. The primer set targeting the N1 region provided more reliable and precise results and for this primer set our method had a limit of detection of 1 viral particle per mg of stool. CONCLUSIONS: Here we describe a sensitive method for detecting SARS-CoV-2 in stool samples. This method can be used to establish the persistence of SARS-CoV-2 in stool and ensure the safety of clinical practices such as FMT.


Subject(s)
COVID-19 Nucleic Acid Testing/methods , COVID-19 , Feces/virology , RNA, Viral/isolation & purification , SARS-CoV-2/isolation & purification , COVID-19/diagnosis , COVID-19/virology , Humans , Limit of Detection
14.
Front Public Health ; 9: 652842, 2021.
Article in English | MEDLINE | ID: covidwho-1389255

ABSTRACT

Background: The viral shedding time (VST) of SARS-CoV-2 mainly determines its transmission and duration of infectiousness. However, it was heterogeneous in the existing studies. Here, we performed a meta-analysis to comprehensively summarize the VST of SARS-CoV-2. Methods: We searched PubMed, Web of Science, MedRxiv, BioRxiv, CNKI, CSTJ, and Wanfang up to October 25, 2020, for studies that reported VSTs of SARS-CoV-2. Pooled estimates and 95% CIs for the VSTs were calculated using log-transformed data. The VSTs in SARS-CoV-2 infections based on different demographic and clinical characteristics, treatments and specimens were stratified by subgroup analysis. Results: A total of 35 studies involving 3,385 participants met the inclusion criteria. The pooled mean VST was 16.8 days (95% CI: 14.8-19.4, I 2 = 99.56%) in SARS-CoV-2 infections. The VST was significantly longer in symptomatic infections (19.7 days, 95% CI: 17.2-22.7, I 2 = 99.34%) than in asymptomatic infections (10.9 days, 95% CI: 8.3-14.3, I 2 = 98.89%) (P < 0.05). The VST was 23.2 days (95% CI: 19.0-28.4, I 2 = 99.24%) in adults, which was significantly longer than that in children (9.9 days, 95% CI: 8.1-12.2, I 2 = 85.74%) (P < 0.05). The VST was significantly longer in persons with chronic diseases (24.2 days, 95% CI: 19.2-30.2, I 2 = 84.07%) than in those without chronic diseases (11.5 days, 95% CI: 5.3-25.0, I 2 = 82.11%) (P < 0.05). Persons receiving corticosteroid treatment (28.3 days, 95% CI: 25.6-31.2, I 2 = 0.00%) had a longer VST than those without corticosteroid treatment (16.2 days, 95% CI: 11.5-22.5, I 2 = 92.27%) (P = 0.06). The VST was significantly longer in stool specimens (30.3 days, 95% CI: 23.1-39.2, I 2 = 92.09%) than in respiratory tract specimens (17.5 days, 95% CI: 14.9-20.6, I 2 = 99.67%) (P < 0.05). Conclusions: A longer VST was found in symptomatic infections, infected adults, persons with chronic diseases, and stool specimens.


Subject(s)
COVID-19/virology , SARS-CoV-2/physiology , Virus Shedding , Adrenal Cortex Hormones/therapeutic use , Adult , Asymptomatic Infections , Child , Comorbidity , Feces/virology , Humans
15.
Virol J ; 18(1): 180, 2021 09 05.
Article in English | MEDLINE | ID: covidwho-1388778

ABSTRACT

BACKGROUND: Covid-19 has the respiratory tract as the main target of infection, and patients present mainly dyspnea, pneumonia, dry cough, and fever. Nevertheless, organs outside the respiratory tract had been reported in recent studies, including the gastrointestinal tract and liver. The host innate immune system recognizes pathogen-associated molecular patterns (PAMPs) through their pattern recognition receptor (PRRs). Toll-like receptor 7 (TLR-7) is a pattern recognition receptor recognizing ssRNA (SARS-CoV-2 is an ssRNA). Polymorphisms are characterized by two or more alternative forms of a distinct phenotype in the same population. Polymorphisms in tlrs genes can negatively influence the immune response to infectious diseases. There are several references in the literature to non-synonymous single nucleotide (rs) polymorphisms related to several genes. Some of them are important for the innate immunity, as rs 179008 (tlr-7), rs3775291 (tlr3), rs8177374 (tir domain-containing adaptor protein, tirap), rs1024611 (monocyte chemoattractant protein-1, mcp-1) and rs61942233 (2'-5'-oligoadenylate synthase-3, oas-3). CASE PRESENTATION: We identified a 5-year-old-male child with gastrointestinal symptoms and fever presenting acholic stool and jaundice, who was positive for SARS-CoV-2 IgM, IgA, and IgG and presenting the Gln11Leu rs 179008 in tlr-7. The child presented high levels of aspartate aminotransferase, alanine aminotransferase, bilirubin, C-reactive protein, D-dimer, gamma-glutamyl transferase, alkaline phosphatase, and was negative for serological tests for hepatitis A, B, C, E, HIV 1 and 2, herpes virus, cytomegalovirus, Epstein-Barr virus, and negative for RTqPCR for Influenza A and B, RSV and SARS-CoV-2. We also investigated other SNPs in the tlr-3 (rs3775291), tirap (rs8177374), mcp-1 (rs1024611), and oas-3 (rs61942233) genes, and no mutation was detected. After an interview with the child's caregivers, any possible accidental ingestion of drugs or hepatotoxic substances was ruled out. CONCLUSION: To our knowledge, this is the first report of a SARS-CoV-2 caused hepatitis in a male child that has the tlr-7 Gln11Leu rs 179008, which could impair an efficient initial immune response. The knowledge of the patient's immune deficiency could improve the treatment to correct this deficiency with specific medications.


Subject(s)
COVID-19/genetics , COVID-19/virology , Hepatitis, Viral, Human/genetics , Hepatitis, Viral, Human/virology , Toll-Like Receptor 7/genetics , Antibodies, Viral/blood , COVID-19/immunology , Child, Preschool , Epstein-Barr Virus Infections/genetics , Epstein-Barr Virus Infections/virology , Feces/virology , Hepatitis, Viral, Human/immunology , Herpesvirus 4, Human/isolation & purification , Humans , Immunity, Innate , Influenza, Human , Male , Polymorphism, Single Nucleotide , SARS-CoV-2/isolation & purification
16.
Pediatr Allergy Immunol ; 31 Suppl 26: 92-95, 2020 11.
Article in English | MEDLINE | ID: covidwho-1388380

ABSTRACT

Gastrointestinal symptoms are common findings in children with SARS-CoV-2 infection. Diarrhea and vomiting have been reported in about 8%-9% of cases, reaching more than 20% in some studies. Children with gastrointestinal involvement appear to be younger than those without, but the severity of the disease seems to be similar between the two groups of subjects. Fecal shedding in children has been reported in 20%-30% of children and has been observed in both those with and those without overt gastrointestinal involvement. Moreover, prolonged fecal elimination, lasting several days after negativization of real-time polymerase chain reaction assay on respiratory swabs, has been reported with variable frequency in children with SARS-CoV-2 infection. These observations raise the question regarding the possibility of oral-fecal transmission and the possible role of children in spreading the infection, particularly when they appear asymptomatic or with gastrointestinal symptoms but with no respiratory involvement, as well as during their convalescent phase.


Subject(s)
COVID-19/complications , Gastrointestinal Diseases/etiology , SARS-CoV-2 , COVID-19/transmission , Child , Feces/virology , Humans , Virus Shedding
19.
Clin Dermatol ; 38(6): 750-756, 2020.
Article in English | MEDLINE | ID: covidwho-1385291

ABSTRACT

Pemphigus and its variants, viz., vulgaris, foliaceous, vegetans, Ig A pemphigus, paraneoplastic pemphigus and Senear-Usher syndrome are rare autoimmune blistering diseases of the skin and/or mucous membranes. The autoantibodies involved in the pathogenesis of pemphigus against desmoglein result in the breach of the skin and mucosal barrier, which acts as the first line of defence against pathogens. In this paper we underscore the importance of the integumentary system as a shield against the acquisition as well as transmission of SARS-CoV-2 virion. We have also made an attempt to delineate the various treatment modalities available and the viral-drug dynamics involved in choosing the optimum therapeutic modality.


Subject(s)
Adrenal Cortex Hormones/therapeutic use , COVID-19/transmission , Pemphigus/drug therapy , Virus Shedding , Administration, Oral , Adrenal Cortex Hormones/adverse effects , COVID-19/complications , Feces/virology , Humans , Intestinal Mucosa/drug effects , Intestinal Mucosa/virology , Mouth Mucosa/virology , Pemphigus/complications , Risk Factors , SARS-CoV-2 , Virus Shedding/drug effects
20.
Pan Afr Med J ; 39: 147, 2021.
Article in English | MEDLINE | ID: covidwho-1377121

ABSTRACT

Introduction: the World health organisation (WHO) African Region reported the first confirmed COVID-19 case caused by the SARS-CoV-2 on 25th February 2020, and the first case for the East Southern Africa (ESA) sub-region was on 5th March 2020. Almost all countries in the ESA sub region implemented the WHO-recommended preventive measures variably after the notification of community transmission of the COVID-19 disease. This resulted in the disruption of the outpatient, immunization surveillance, and the related supply chain activities. Methods: a comparative analysis study design of secondary acute flaccid paralysis (AFP) surveillance data received from the East and Southern Africa sub-region countries to evaluate the effect of the COVID-19 pandemic in the AFP field surveillance for the same time period of March to December 2019 and 2020. Results: we observed that 52.4% of second stool samples were received in the laboratory within 72 hours from March to December 2019, and only 48.1% in the same period of 2020. A 4.3% decline with a p-value of <0.0001 (95% CI, ranges from 2.326% to 6.269%). Similarly, we noted a 4.7% decline in the number of reported AFP cases in the ESA sub-region for March to December 2020 compared to the same period in 2019, a p-value of less than 0.001 (95% CI ranges from 2.785 to 6.614). For the percentage of stool adequacy, we observed a 3.37% decline for April in 2020 compared to April 2019 with a p-value of less than 0.001 (95% CI ranges from 2.059 to 4.690). Conclusion: we observed a decline in the core AFP surveillance (non polio) NP-AFP rate, and percentage of stool adequacy in countries severely affected by the COVID-19 disease. These countries implemented stringent transmission prevention measures such as lock-down and international transportation restrictions.


Subject(s)
COVID-19 , Central Nervous System Viral Diseases/diagnosis , Feces/virology , Myelitis/diagnosis , Neuromuscular Diseases/diagnosis , Population Surveillance/methods , Adolescent , Africa, Eastern/epidemiology , Africa, Southern/epidemiology , Central Nervous System Viral Diseases/epidemiology , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Myelitis/epidemiology , Neuromuscular Diseases/epidemiology
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