ABSTRACT
Objective:To analyze the composition and functional characteristics of the intestinal microflora in novel coronavirus pneumonia patients with arterial stiffness, in order to provide empirical evidence for rational use of intestinal microecological modulators.Methods:Patients with novel coronavirus pneumonia admitted to our hospital between April 5 and April 19, 2020 were enrolled as research subjects.They were divided into the stiffness group and the control group according to the results of arterial stiffness.Stool samples were collected within 7 days of admission.Intestinal flora DNA was analyzed and entered into a database, shotgun metagenomic sequencing was performed, and bioinformatics analysis was conducted based on sequencing results.Results:A total of 16 patients with novel coronavirus pneumonia were included in this study, including 7 in the stiffness group and 9 in the control group.Brachial-ankle pulse wave velocity and blood pressure were higher in the stiffness group than in the control group( P<0.05). Beta diversity analysis at the phylum level showed that there were significant differences in the composition of the intestinal flora between the two groups( P<0.05). Patients with arterial stiffness had a lower relative abundance than the controls in fecal Holdemanella, Mitsuokella, Deinococcus, Lachnospira, Turicibacter, Butyrivibrio, Sporomusa, and Halanaerobium, species associated with the production of short-chain fatty acids(SCFAs), regulation of energy metabolism, anti-radiation, anti-oxidative stress and anti-inflammatory effects.The Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis showed that the intestinal flora in the control group was mainly involved in the pathways of lipopolysaccharide biosynthesis, metabolism of SCFA and other amino acids, and membrane transport, while the intestinal flora in the stiffness group was mostly concerned with the pathways of amino acid metabolism and DNA damage repair. Conclusions:The use of microbial agents capable of increasing short-chain fatty acids in patients with novel caronavirus infection may contribute to the restoration of intestinal flora homeostasis.
ABSTRACT
Objective To establish human metapneumovirus(hMPV)nucleotide rapid,specific and sensitive gene chip detec-tion method,and provide effective diagnostic methods for hMPV detection and diagnosis in shenzhen area.Methods The flu-orescence PCR primer of hMPV was designed for the highly conservative regional gene sequence of hMPV virus.Application of array probe design software designer 4.20 hMPV oligonucleotide detection probe design,oligonucleotide probe sample points to aldehyde slides on the preparation of hMPV gene chip,and parallel compared with conventional reverse tran-scriptase-polymerase chain reaction(RT-PCR),the sensitivity,specificity and repeatability,and was used to evaluate the clinical applicability of the samples.Results hMPV gene chip method to detect hMPV specificity was 96.23%(230/239), the sensitivity was 2.0×101/μl,linear range was 2.0×101~2.0×107/μl,and the repeatability was good.Initial tests in shenzhen area 300 clinical specimens on a nasopharyngeal swab,gene chip method detection rate was 20.3%(61/300),sig-nificantly higher than the conventional RT-PCR method 9.7%(29/300),the difference was statistically significant between the sensitivity of two methods(χ2=39.205,P<0.05).The results were consistent in two methods(kappa=0.360 7).Con-clusion Established hMPV microarray assay,gene chip method to detect hMPV has high specificity and sensitivity,wide linear range and detection.The popularity of hMPV monitoring for laboratory provides a new detecting technology and early diagnosis.
ABSTRACT
Objective To investigate the prevalence of human metapneumovirus (HMPV)infection in patients with respiratory infection in Fuzhou area and compare their epidemic features and clinical characteristics with those of infected with respiratory syncytial virus (RSV). Methods A total of 153 sputum or pharyngeal swab samples from patients with respiratory tract infection were collected in Fujian Provincial Hospital in consecutive winter and spring seasons from 2005 to 2007. HMPV was detected by nested reverse transcriptase-polymerase chain reaction (RT-PCR) and RSV was tested by RT-PCR. Parts of PCR products were sequenced and analyzed using DNAMAN software. The clinical symptoms, signs and epidemiology of the respiratory tract infections caused by HMPV and RSV were compared. Results In the 153 specimens, 32 (20.9%) were positive for H MPV, 26 (17.0%) were positive for RSV, and 8 were both HMPV and RSV positive. Nucleotide sequences of three 432-bp PCR products were 100% identical and submitted to GenBank (the accession No. DQ887758).Phylogenetic tree analysis of nucleotide sequences revealed that the three isolates clustered in HMPV belonged to genotype A with part of mutation. Twenty-six samples (26. 7%) were HMPV positive from Dec 2005 to Apr 2006 and 6 (10.7%) were positive from Dec 2006 to Apr 2007. The RSV detection rate was opposite of HMPV. The mean age of RSV infection in children was (2.65±2.65)years old and HMPV infection was (4.58 ±3.35) years old. The main clinical manifestations of both RSV and HMPV infections were cough, sore throat and fever. Conclusions Both HMPV and RSV are the major pathogens of respiratory tract infection in Fuzhou area and HMPV and RSV coinfection could be detected. HMPV infected children are older. The clinical features of HMPV and RSV infections are similar. Single genotype of HMPV is detected in Fuzhou area in this study.