Comparison of Throat and Nasopharyngeal Swab Specimens for Molecular Diagnosis of Mycoplasma pneumoniae

A loop-mediated isothermal amplification (LAMP) assay for the rapid detection of Mycoplasma pneumoniae was developed and evaluated. The assay specifically amplified only M. pneumoniae sequences, and no cross-reactivity was observed for other Mycoplasma species or respiratory bacterial species. The detection limit for this assay was found to be 2 × 102 copies, corresponding to 2–20 colour changing units of M. pneumoniae in 1 h, as observed in a real-time turbidimeter and electrophoretic analysis. The accuracy of the LAMP reaction was confirmed by restriction endonuclease analysis as well as direct sequencing of the amplified product. The assay was applied to 95 nasopharyngeal swab samples collected from patients or from healthy individuals, and compared to a real-time PCR assay in-house. A concordance of 100 % was observed between the two assays. The LAMP assay is easy to perform, shows a rapid reaction and is inexpensive. It may therefore be applied in the routine diagnosis of M. pneumoniae infection in the clinical laboratory.

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Mycoplasma pneumoniae Infections in a Tertiary Care Hospital

Journal of Advances in Medicine and Medical Research ◽

10.9734/jammr/2021/v33i2131128 ◽

2021 ◽

pp. 8-14

Author(s):

Harun Agca

Keyword(s):

Mycoplasma Pneumoniae ◽

Nasal Congestion ◽

Tertiary Care ◽

Hospitalized Patients ◽

Nasopharyngeal Swab ◽

Nasal Discharge ◽

Atypical Pneumonia ◽

Care Hospital ◽

Age Group ◽

Tract Infections

Background: Mycoplasma pneumoniae is a common causative agent of community-acquired and atypical pneumonia. M. pneumoniae-associated respiratory tract infections are most often mild and self-limiting in nature, and severe courses of disease are not common. Aims and Objectives: We aimed to analyze respiratory samples of hospitalized patients by real-time (RT) PCR for M. pneumoniae infections. Methodology: In this study, hospitalized patients with respiratory symptoms including nasal congestion, rhinorrhea, nasal discharge, wheezing, sore throat and cough, were tested for Mycoplasma pneumoniae between April 2018 and March 2020 and the results of these patients were analyzed retrospectively. Results: We retrospectively investigated the results of 1245 nasopharyngeal swab samples. Records of 693 (55.7%) male and 552 (44.3%) female patients between 0-93 years were investigated.All of the patients were below 26 years. There were 37 M. pneumoniae positive patients and the highest M. pneumoniae positivity was in 2-5 age group, with 16 children. M. pneumoniae positivity rate was highest in the 0-1 age group with 8.3%. There were 19 co-infections with viruses and rhino/enterovirus (n=12) was the most frequent accompanying virus. Conclusion: M. pneumonia infections are found in about 3% of respiratory samples. M. pneumoniae is usually a part of co-infections accompanied by bacteria and viruses. Patients with respiratory infection symptoms should be diagnosed with multiplex rapid PCR panels including M. pneumoniae.

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Molecular diagnosis of adenovirus, mycoplasma pneumoniae and Chlamydia pneumoniae infection in hospitalized children

10.5353/th_b3197212 ◽

2004 ◽

Author(s):

Chi-kit, Patrick Pun

Keyword(s):

Mycoplasma Pneumoniae ◽

Molecular Diagnosis ◽

Chlamydia Pneumoniae ◽

Hospitalized Children

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Molecular diagnosis of NPC-detecting LMP-1 and EBNA by nasopharyngeal swab

Otolaryngology ◽

10.1016/s0194-5998(03)01235-x ◽

2003 ◽

Vol 129 (2) ◽

pp. P198

Author(s):

S Hao

Keyword(s):

Molecular Diagnosis ◽

Nasopharyngeal Swab

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Molecular Diagnosis of Nasopharyngeal Carcinoma: Detecting LMP-1 and EBNA by Nasopharyngeal Swab

Yearbook of Pathology and Laboratory Medicine ◽

10.1016/s1077-9108(08)70132-4 ◽

2006 ◽

Vol 2006 ◽

pp. 175-176

Author(s):

A.V. Parwani

Keyword(s):

Nasopharyngeal Carcinoma ◽

Molecular Diagnosis ◽

Nasopharyngeal Swab

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Molecular diagnosis of SARS-CoV-2 in seminal fluid

Journal of Endocrinological Investigation ◽

10.1007/s40618-021-01580-x ◽

2021 ◽

Author(s):

D. Paoli ◽

F. Pallotti ◽

G. Nigro ◽

L. Mazzuti ◽

M. N. Hirsch ◽

...

Keyword(s):

Molecular Diagnosis ◽

Seminal Plasma ◽

Reproductive Medicine ◽

Seminal Fluid ◽

Rna Extraction ◽

Nasopharyngeal Swab ◽

Control Panel ◽

Rt Pcr ◽

Sperm Bank ◽

University Of Rome

Abstract Purpose Due to relevant repercussions on reproductive medicine, we aimed to evaluate feasibility of RT-PCR as a detection method of SARS-CoV-2 RNA in seminal fluid. Methods A qualitative determination of the RT-PCR assays in semen was performed through different approaches: (1) efficiency of RNA extraction from sperm and seminal plasma was determined using PRM1 and PRM2 mRNA and a heterologous system as control; (2) samples obtained by diluting viral preparation from a SARS-CoV-2 panel (virus cultured in Vero E6 cell lines) were tested; (3) viral presence in different fractions of seminal fluid (whole sample, seminal plasma and post-centrifugation pellet) was evaluated. Semen samples from mild and recovered COVID-19 subjects were collected by patients referring to the Infectious Disease Department of the Policlinico Umberto I Hospital - “Sapienza” University of Rome. Control subjects were recruited at the Laboratory of Seminology-Sperm Bank “Loredana Gandini'' of the same hospital. Results The control panel using viral preparations diluted in saline and seminal fluid showed the capability to detect viral RNA presence with Ct values depending on the initial viral concentration. All tested semen samples were negative for SARS-CoV-2, regardless of the nasopharyngeal swab result or seminal fluid fraction. Conclusion These preliminary data show that RT-PCR for SARS-CoV-2 RNA testing appears to be a feasible method for the molecular diagnosis of SARS-CoV-2 in seminal fluid, supported by results of the control panel. The ability to detect SARS-CoV-2 in semen is extremely important for reproductive medicine, especially in assisted reproductive technology and sperm cryopreservation.

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Identification of Coinfections by Viral and Bacterial Pathogens in COVID-19 Hospitalized Patients in Peru: Molecular Diagnosis and Clinical Characteristics

Antibiotics ◽

10.3390/antibiotics10111358 ◽

2021 ◽

Vol 10 (11) ◽

pp. 1358

Author(s):

Giancarlo Pérez-Lazo ◽

Wilmer Silva-Caso ◽

Juana del Valle-Mendoza ◽

Adriana Morales-Moreno ◽

José Ballena-López ◽

...

Keyword(s):

Hospital Admission ◽

Mycoplasma Pneumoniae ◽

Molecular Diagnosis ◽

Clinical Characteristics ◽

Chlamydia Pneumoniae ◽

Severe Pneumonia ◽

Hospitalized Patients ◽

Respiratory Pathogens ◽

Ministry Of Health ◽

The Impact

The impact of respiratory coinfections in COVID-19 is still not well understood despite the growing evidence that consider coinfections greater than expected. A total of 295 patients older than 18 years of age, hospitalized with a confirmed diagnosis of moderate/severe pneumonia due to SARS-CoV-2 infection (according to definitions established by the Ministry of Health of Peru) were enrolled during the study period. A coinfection with one or more respiratory pathogens was detected in 154 (52.2%) patients at hospital admission. The most common coinfections were Mycoplasma pneumoniae (28.1%), Chlamydia pneumoniae (8.8%) and with both bacteria (11.5%); followed by Adenovirus (1.7%), Mycoplasma pneumoniae/Adenovirus (0.7%), Chlamydia pneumoniae/Adenovirus (0.7%), RSV-B/Chlamydia pneumoniae (0.3%) and Mycoplasma pneumoniae/Chlamydia pneumoniae/Adenovirus (0.3%). Expectoration was less frequent in coinfected individuals compared to non-coinfected (5.8% vs. 12.8%). Sepsis was more frequent among coinfected patients than non-coinfected individuals (33.1% vs. 20.6%) and 41% of the patients who received macrolides empirically were PCR-positive for Mycoplasma pneumoniae and Chlamydia pneumoniae.

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Molecular Characterization of Mycoplasma pneumoniae Infections in Two Rural Populations of Thailand from 2009 to 2012

Journal of Clinical Microbiology ◽

10.1128/jcm.00350-17 ◽

2017 ◽

Vol 55 (7) ◽

pp. 2222-2233 ◽

Cited By ~ 10

Author(s):

Toni Whistler ◽

Pongpun Sawatwong ◽

Maureen H. Diaz ◽

Alvaro J. Benitez ◽

Bernard J. Wolff ◽

...

Keyword(s):

Molecular Characterization ◽

Mycoplasma Pneumoniae ◽

Variable Number Tandem Repeat ◽

Population Based ◽

Variable Number ◽

23S Rrna ◽

Nasopharyngeal Swab ◽

Urban Centers ◽

Content Type ◽

Adhesion Protein

ABSTRACTStudies onMycoplasma pneumoniaein Thailand have focused on urban centers and have not included molecular characterization. In an attempt to provide a more comprehensive understanding of this organism, we conducted a systematic random sampling to identify 3,000 nasopharyngeal swab specimens collected from January 2009 through July 2012 during population-based surveillance for influenza-like illness in two rural provinces.M. pneumoniaewas detected by real-time PCR in 175 (5.8%) specimens. Genotyping was performed using the major adhesion protein (P1) and multilocus variable-number tandem-repeat analysis (MLVA). Of the 157 specimens typed, 97 were P1 type 1 and 60 were P1 type 2. Six different MLVA profiles were identified in 149 specimens, with 4/5/7/2 (40%) and 3/5/6/2 (26%) predominating. There was no discrete seasonality toM. pneumoniaeinfections. Examination of the 23S rRNA sequence for known polymorphisms conferring macrolide resistance revealed that all 141 tested to possess the genotype associated with macrolide susceptibility.

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