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      Molecular Characterization of Mycoplasma pneumoniae Isolates in the United States from 2012 to 2018

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          Abstract

          Mycoplasma pneumoniae is a major cause of community-acquired pneumonia. There are limited data in the United States on the molecular epidemiological characteristics of M. pneumoniae. We collected 446 M. pneumoniae-positive specimens from 9 states between August 2012 and October 2018. Culture, antimicrobial susceptibility testing, P1 subtyping, and multilocus VNTR (variable-number tandem repeats) analysis (MLVA) were performed to characterize the isolates.

          ABSTRACT

          Mycoplasma pneumoniae is a major cause of community-acquired pneumonia. There are limited data in the United States on the molecular epidemiological characteristics of M. pneumoniae. We collected 446 M. pneumoniae-positive specimens from 9 states between August 2012 and October 2018. Culture, antimicrobial susceptibility testing, P1 subtyping, and multilocus VNTR (variable-number tandem repeats) analysis (MLVA) were performed to characterize the isolates. Macrolide-resistant M. pneumoniae (MRMp) was detected in 37 (8.3%) specimens. P1 subtype 2 (P1-2) was the predominant P1 subtype (59.8%). P1 subtype distribution did not change significantly chronologically or geographically. The macrolide resistance rate in P1 subtype 1 (P1-1) samples was significantly higher than that in P1-2 (12.9% versus 5.5%). Six P1-2 variants were identified, including two novel types, and variant 2c was predominant (64.6%). P1-2 variants were distributed significantly differently among geographic regions. Classical P1-2 was more frequent in lower respiratory tract specimens and had longer p1 trinucleotide repeats. Classical P1-2 was most common in MRMp (35.7%), while variant 2c was most common in macrolide-susceptible M. pneumoniae (67.5%). Fifteen MLVA types were identified; 3-5-6-2 (41.7%), 4-5-7-2 (35.3%), and 3-6-6-2 (16.6%) were the major types, and four MLVA clusters were delineated. The distribution of MLVA types varied significantly over time and geographic location. The predominant MLVA type switched from 4-5-7-2 to 3-5-6-2 in 2015. MLVA type was associated with P1 subtypes and P1-2 variant types but not with macrolide resistance. To investigate the M. pneumoniae genotype shift and its impact on clinical presentations, additional surveillance programs targeting more diverse populations and prolonged sampling times are required.

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          Most cited references58

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          Community-acquired pneumonia requiring hospitalization among U.S. children.

          Incidence estimates of hospitalizations for community-acquired pneumonia among children in the United States that are based on prospective data collection are limited. Updated estimates of pneumonia that has been confirmed radiographically and with the use of current laboratory diagnostic tests are needed.
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            Epidemiology of Mycoplasma pneumoniae Infections in Japan and Therapeutic Strategies for Macrolide-Resistant M. pneumoniae

            Pneumonia caused by Mycoplasma pneumoniae (M. pneumoniae pneumonia) is a major cause of community-acquired pneumonia worldwide. The surveillance of M. pneumoniae pneumonia is important for etiological and epidemiological studies of acute respiratory infections. In Japan, nation-wide surveillance of M. pneumoniae pneumonia has been conducted as a part of the National Epidemiological Surveillance of Infectious Diseases (NESID) program. This surveillance started in 1981, and significant increases in the numbers of M. pneumoniae pneumonia patients were noted in 1984, 1988, 2006, 2010, 2011, 2012, and 2015. The epidemics in 2011 and 2012 were particularly widespread and motivated researchers to conduct detailed epidemiological studies, including genotyping and drug resistance analyses of M. pneumoniae isolates. The genotyping studies based on the p1 gene sequence suggested that the p1 gene type 1 lineage has been dominant in Japan since 2003, including the epidemic period during 2011–2012. However, more detailed p1 typing analysis is required to determine whether the type 2 lineages become more relevant after the dominance of the type 1 lineage. There has been extensive research interest in implications of the p1 gene types on the epidemiology of M. pneumoniae infections. Serological characterizations of sera from patients have provided a glimpse into these associations, showing the presence of type specific antibody in the patient sera. Another important epidemiological issue of M. pneumoniae pneumonia is the emergence of macrolide-resistant M. pneumoniae (MRMP). MRMPs were noted among clinical isolates in Japan after 2000. At present, the isolation rate of MRMPs from pediatric patients is estimated at 50–90% in Japan, depending on the specific location. In view of the situation, Japanese societies have issued guiding principles for treating M. pneumoniae pneumonia. In these guiding principles, macrolides are still recommended as the first-line drug, however, if the fever does not subside in 48–72 h from first-line drug administration, a change of antibiotics to second-line drugs is recommended.
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              Development of multiple-locus variable-number tandem-repeat analysis for molecular typing of Mycoplasma pneumoniae.

              In this study we report on the development of a multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) method for the molecular typing of Mycoplasma pneumoniae. The genomic content of M. pneumoniae M129 was analyzed for VNTRs, and 5 of the 17 VNTRs identified were selected for use in an MLVA assay. The method was based on a GeneScan analysis of VNTR loci labeled with fluorescent dyes by multiplex PCR and capillary electrophoresis. This approach was applied to a collection of 265 isolates from various European countries, Japan, and Tunisia; and 26 distinct VNTR types were found. The VNTR assay was compared to the P1 adhesin PCR-restriction fragment length polymorphism (RFLP) typing method and showed a far better resolution than the P1 PCR-RFLP method. The discriminatory power of MLVA (Hunter-Gaston diversity index [HGDI], 0.915) for the 265 isolates was significantly higher than that of the P1 PCR-RFLP method (HGDI, 0.511). However, there was a correlation between the typing results obtained by MLVA and the P1 gene PCR-RFLP method. The potential value of MLVA of M. pneumoniae as an epidemiological tool is discussed, and the use of the VNTR markers in further investigations of the potential use of MLVA in outbreaks of M. pneumoniae infections is proposed.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                J Clin Microbiol
                J. Clin. Microbiol
                jcm
                jcm
                JCM
                Journal of Clinical Microbiology
                American Society for Microbiology (1752 N St., N.W., Washington, DC )
                0095-1137
                1098-660X
                22 September 2020
                October 2020
                22 September 2020
                : 58
                : 10
                : e00710-20
                Affiliations
                [a ]Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA
                [b ]Department of Pathology, University of Alabama at Birmingham, Birmingham, Alabama, USA
                [c ]Department of Pediatrics, University of Alabama at Birmingham, Birmingham, Alabama, USA
                [d ]Department of Laboratory Medicine and Pathology, Seattle Children’s Hospital, Seattle, Washington, USA
                [e ]Children’s Mercy Hospital, Kansas City, Missouri, USA
                [f ]Memorial Sloan Kettering Cancer Center, Weill Medical College of Cornell University, New York, New York, USA
                [g ]Ann & Robert H. Lurie Children’s Hospital of Chicago, Chicago, Illinois, USA
                [h ]Department of Pathology and Laboratory Medicine, Children’s Hospital Los Angeles, Keck School of Medicine, University of Southern California, Los Angeles, California, USA
                [i ]Department of Pathology, Hackensack University Medical Center, Hackensack, New Jersey, USA
                [j ]Departments of Pediatrics, University of Texas Health Science Center, San Antonio, Texas, USA
                [k ]Departments of Pathology, University of Texas Health Science Center, San Antonio, Texas, USA
                Carter BloodCare & Baylor University Medical Center
                Author notes
                Address correspondence to L. Xiao, lixiao@ 123456uabmc.edu .

                Citation Xiao L, Ratliff AE, Crabb DM, Mixon E, Qin X, Selvarangan R, Tang Y-W, Zheng X, Dien Bard J, Hong T, Prichard M, Brooks E, Dallas S, Duffy LB, Fowler KB, Atkinson TP, Waites KB. 2020. Molecular characterization of Mycoplasma pneumoniae isolates in the United States from 2012 to 2018. J Clin Microbiol 58:e00710-20. https://doi.org/10.1128/JCM.00710-20.

                Author information
                https://orcid.org/0000-0003-0188-4023
                https://orcid.org/0000-0003-4897-9943
                https://orcid.org/0000-0003-0524-9473
                https://orcid.org/0000-0002-8775-765X
                Article
                00710-20
                10.1128/JCM.00710-20
                7512161
                32817226
                5cdf41d7-036b-41fe-abd8-4bdc1cf1afb2
                Copyright © 2020 Xiao et al.

                This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

                History
                : 10 April 2020
                : 13 May 2020
                : 6 August 2020
                Page count
                supplementary-material: 1, Figures: 6, Tables: 5, Equations: 0, References: 69, Pages: 16, Words: 10716
                Funding
                Funded by: HHS | NIH | National Cancer Institute (NCI), https://doi.org/10.13039/100000054;
                Award ID: CA008748
                Award Recipient :
                Funded by: HHS | Centers for Disease Control and Prevention (CDC), https://doi.org/10.13039/100000030;
                Award ID: 200-2017-96217
                Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient :
                Categories
                Epidemiology
                Custom metadata
                October 2020

                Microbiology & Virology
                mycoplasma pneumoniae,genotype,macrolide resistance,p1,variant,mlva
                Microbiology & Virology
                mycoplasma pneumoniae, genotype, macrolide resistance, p1, variant, mlva

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