Evaluation of a novel immunochromatographic assay using silver amplification technology for detection of Mycoplasma pneumoniae from throat swab samples in pediatric patients

Abstract Objectives Mycoplasma pneumoniae is one of the common causative pathogens of community-acquired respiratory tract infections mainly in children and young adults. Rapid and accurate diagnostic techniques for identifying the causative pathogen would be useful for initiating treatment with an appropriate antibiotic. The purpose of the present study was to evaluate the sensitivity and specificity of a novel immunochromatographic assay using silver amplification technology using FUJI DRI-CHEM IMMUNO AG2 and FUJI DRI-CHEM IMMUNO AG cartridge Myco (FUJIFILM Co., Tokyo, Japan) for detection of M. pneumoniae. Methods Throat swab samples were collected from 170 pediatric patients who were diagnosed with bronchitis or pneumonia. The silver amplification immunochromatographic (SAI) assay was performed using these samples and the results were compared with those of real-time PCR. The time required for the SAI assay is approximately 20 min (5 min for sample preparation and 15 min for waiting time after starting the assay). Results The sensitivity and specificity of the SAI assay for detection of M. pneumoniae were 85.2 and 99.1%, respectively, and the assay showed positive and negative predictive values of 98.1 and 92.3%, respectively, compared with the results of real-time PCR. The diagnostic accuracy was 94.1%. Conclusions FUJI DRI-CHEM IMMUNO AG2 and FUJI DRI-CHEM IMMUNO AG cartridge Myco are appropriate for clinical use. The optimal timing of this assay is five days or more after the onset of M. pneumoniae infection. However, PCR or other molecular methods are superior, especially with regard to sensitivity and negative predictive value.

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Evaluation of a Novel Immunochromatographic Assay for Detection of Human Adenoviruses from Throat Swab Samples Compared with Real-time PCR

Clinical Laboratory ◽

10.7754/clin.lab.2018.180420 ◽

2018 ◽

Vol 64 (10/2018) ◽

Author(s):

Nobuhisa Ishiguro ◽

Miki Kaiho ◽

Hideaki Kikuta ◽

Takehiro Togashi ◽

Daisuke Sato ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Throat Swab ◽

Immunochromatographic Assay ◽

Human Adenoviruses

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Clinical spectrum and diagnostic yields of Mycoplasma pneumoniae as a causative agent of community-acquired pneumonia

Journal of Laboratory Physicians ◽

10.4103/jlp.jlp_62_17 ◽

2018 ◽

Vol 10 (01) ◽

pp. 044-049 ◽

Cited By ~ 2

Author(s):

Saroj Dash ◽

Rama Chaudhry ◽

Benu Dhawan ◽

Aparajit Ballav Dey ◽

Sushil Kumar Kabra ◽

...

Keyword(s):

Real Time ◽

Mycoplasma Pneumoniae ◽

Real Time Pcr ◽

Clinical Signs ◽

Signs And Symptoms ◽

Community Acquired Pneumonia ◽

Diagnostic Methods ◽

Conventional Pcr ◽

Clinical Signs And Symptoms ◽

Tract Infections

Abstract INTRODUCTION: Infection with Mycoplasma pneumoniae (M. pneumonia) occurs worldwide which accounts for 15%–20% of cases of community-acquired pneumonia and indistinguishable clinically from other infectious causes of pneumonia. AIM: The aim of this study was to evaluate the real-time polymerase chain reaction (PCR) and to correlate it with other diagnostic methods such as culture, serology (ELISA), and conventional PCR along with the clinical signs and symptoms produced by M. pneumonia. MATERIALS AND METHODS: A total of 130 patients of all age groups presenting with clinical features of lower respiratory tract infections were enrolled over a period of 1 year and 2 months in a tertiary care hospital in Delhi. M. pneumoniae in throat swab samples was detected by real-time PCR, compared with culture, serology, conventional PCR, and clinical signs and symptoms. Univariate analyses were conducted to determine the association of M. pneumoniae infection among different categories of patients. RESULTS: Out of a total of 130 patients, 18 patients (14%) were positive for M. pneumoniae by any test; culture was positive in nine patients (50%), serology (IgM) in eight patients (44.4%), PCR in five patients (27.7%), and real-time PCR was positive in six patients (33.3%). Clinical signs and symptoms were higher in incidence in M. pneumoniae-positive patients. Age-matched healthy controls (30) were included in the study, and all were negative for any diagnostic test performed (P = 0.026). CONCLUSION: It was concluded that combination of M. pneumoniae-specific testing modalities is required for the diagnosis of this etiological agent rather than a single diagnostic method.

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Evaluation of the ELITe InGenius PCR Platform for Detection of Mycoplasma pneumoniae

Journal of Clinical Microbiology ◽

10.1128/jcm.00287-19 ◽

2019 ◽

Vol 57 (6) ◽

Cited By ~ 2

Author(s):

Arthur H. Totten ◽

Sixto M. Leal ◽

Amy E. Ratliff ◽

Li Xiao ◽

Donna M. Crabb ◽

...

Keyword(s):

Real Time ◽

Mycoplasma Pneumoniae ◽

Real Time Pcr ◽

Growth Media ◽

Test Accuracy ◽

Analytical Sensitivity ◽

Pcr Assay ◽

Predictive Values ◽

Content Type ◽

Significant Difference

ABSTRACT Mycoplasma pneumoniae is the leading cause of bacterial community-acquired pneumonia in persons of all ages. Due to the fastidious nature of this bacterium and the necessary specialized growth media, nucleic acid amplification testing is currently the most reliable means for patient diagnostics. Analytical sensitivity, specificity, reproducibility, and clinical performance of the ELITe InGenius automated PCR platform with its MGB Alert M. pneumoniae real-time PCR research use only reagents (ELITechGroup, Inc., Bothell, WA) were compared with those of a laboratory-developed real-time PCR assay targeting repMp1 for detection of M. pneumoniae. The ELITe InGenius PCR assay successfully detected 31 distinct M. pneumoniae clinical isolates and reference strains, and there was no cross-reactivity with other mollicutes, Gram-positive bacteria, or Gram-negative bacteria. In testing 223 clinical samples, the ELITe InGenius PCR showed 95.79% and 99.22% positive and negative agreement with the repMp1 assay, respectively. Additionally, the ELITech platform showed 98.91% positive and 96.95% negative predictive values, and there was no significant difference detected between the two assays (McNemar’s test, P = 0.375). The ELITe InGenius PCR assay limit of detection was 0.16 CFU/PCR test or 4.16 genome copies (GCs)/test. Accuracy, instrument ease-of-use, and decreased hands-on time make the ELITe InGenius platform suitable for detection of M. pneumoniae directly from clinical specimens.

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Molecular Diagnosis of Tubercular Lymphadenopathy from Fine-Needle Aspirates in Pediatric Patients

Acta Cytologica ◽

10.1159/000475832 ◽

2017 ◽

Vol 61 (3) ◽

pp. 173-178 ◽

Cited By ~ 2

Author(s):

Vivek Gupta ◽

Arvind Bhake

Keyword(s):

Real Time ◽

Sensitivity And Specificity ◽

Real Time Pcr ◽

Pediatric Patients ◽

Strong Association ◽

Immunological Response ◽

Lymphoid Hyperplasia ◽

Reactive Lymphoid Hyperplasia ◽

Cross Sectional ◽

Fine Needle

Objectives: The diagnosis of peripheral tubercular lymphadenopathy (TBLN) in pediatric patients is often a challenge because features evident on fine-needle aspiration cytology (FNAC) or tissue biopsy can be deceptive for the reason that they result from an immunological response. This study aimed to evaluate polymerase chain reaction (PCR) for Mycobacterium tuberculosis complex (MTBC) in pediatric patients under clinical suspicion for TBLN and to assess its role in the evaluation of cases cytodiagnosed as reactive lymphoid hyperplasia. Methods: This was a cross-sectional study conducted on 45 pediatric patients clinically suspected and unsuspected for TBLN. FNAC, culture on Löwenstein-Jensen medium, and real-time PCR were performed. Comparative values with reference to the culture were calculated. Results: Cytology had a sensitivity and specificity of 38.5 and 87.5%, respectively. Real-time PCR had a sensitivity and specificity of 84.6 and 81.3%, respectively. Of the 32 cases with a cytodiagnosis of reactive lymphoid hyperplasia, 53% were positive both on PCR and culture for M. tuberculosis; the φ value of 0.93 demonstrated a strong association between these 2 methods. Conclusion: Real-time PCR is useful in detecting MTBC in pediatric patients, and it also helps in the diagnosis of cases missed on FNAC.

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Evaluation of new immunochromatographic assay kit for adenovirus detection in throat swab: Comparison with culture and real-time PCR results

Journal of Infection and Chemotherapy ◽

10.1016/j.jiac.2014.01.005 ◽

2014 ◽

Vol 20 (5) ◽

pp. 303-306 ◽

Cited By ~ 5

Author(s):

Miyuki Morozumi ◽

Hideaki Shimizu ◽

Yuki Matsushima ◽

Keiko Mitamura ◽

Takeshi Tajima ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Throat Swab ◽

Immunochromatographic Assay

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Diagnostic Accuracy of a Noninvasive Test for Detection of Helicobacter pylori and Resistance to Clarithromycin in Stool by the Amplidiag H. pylori+ClariR Real-Time PCR Assay

Journal of Clinical Microbiology ◽

10.1128/jcm.01787-19 ◽

2020 ◽

Vol 58 (4) ◽

Cited By ~ 6

Author(s):

Maxime Pichon ◽

Benoit Pichard ◽

Thierry Barrioz ◽

Chloé Plouzeau ◽

Vincent Croquet ◽

...

Keyword(s):

Helicobacter Pylori ◽

Real Time ◽

Sensitivity And Specificity ◽

Real Time Pcr ◽

Rrna Genes ◽

23S Rrna ◽

Noninvasive Test ◽

Predictive Values ◽

Content Type ◽

H Pylori

ABSTRACT The noninvasive detection of Helicobacter pylori and its resistance to clarithromycin could revolutionize the management of H. pylori-infected patients by tailoring eradication treatment without any need for endoscopy when histology is not necessary. Several real-time PCR tests performed on stools have been proposed, but their performances were either poor or they were tested on too few patients to be properly evaluated. We conducted a prospective, multicenter study including 1,200 adult patients who were addressed for gastroduodenal endoscopy with gastric biopsies and who were naive for eradication treatment in order to evaluate the performance of the Amplidiag H. pylori+ClariR assay recently developed by Mobidiag (Espoo, Finland). The results of the Amplidiag H. pylori+ClariR assay performed on DNA from stools (automatic extraction with the EasyMag system [bioMérieux]) were compared with those of culture/Etest and quadruplex real-time PCRs performed on two gastric biopsy samples (from the antrum and corpus) to detect the H. pylori glmM gene and mutations in the 23S rRNA genes conferring clarithromycin resistance. The sensitivity and specificity of the detection of H. pylori were 96.3% (95% confidence interval [CI], 92 to 98%) and 98.7% (95% CI, 97 to 99%), respectively. The positive and negative predictive values were evaluated to be 92.2% (95% CI, 92 to 98%) and 99.3% (95% CI, 98 to 99%), respectively. In this cohort, 160 patients (14.7%) were found to be infected (positive by culture and/or PCR). The sensitivity and specificity for detecting resistance to clarithromycin were 100% (95% CI, 88 to 100%) and 98.4% (95% CI, 94 to 99%), respectively.

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Evaluation of significant bacteriuria in pregnant women using quantitative real-time PCR

Journal of obstetrics and women s diseases ◽

10.17816/jowd65450-56 ◽

2016 ◽

Vol 65 (4) ◽

pp. 50-56

Author(s):

Tatyana A. Khusnutdinova ◽

Elena V Shipitsina ◽

Yulia A. Savochkina ◽

Olga Yu. Timoshina ◽

Elena V. Rybina ◽

...

Keyword(s):

Pregnant Women ◽

Real Time ◽

Sensitivity And Specificity ◽

Real Time Pcr ◽

Urine Culture ◽

Urine Samples ◽

Quantitative Real Time Pcr ◽

Diagnostic Characteristics ◽

Significant Bacteriuria ◽

Tract Infections

Introduction. Urinary tract infections are the most common infections in obstetrics and gynecology. Bacteriological method to investigate urine is laborious and time-consuming, therefore development of accurate and rapid methods for the detection of significant bacteriuria is important. Objective. Evaluation of quantitative real-time PCR based approach for the detection of significant bacteriuria in pregnant women. Material and methods. A retrospective investigation of mid-stream urine samples obtained from pregnant women was performed. Urine culture was performed using quantitative method, and a case was considered as significant bacteriuria if ≥ 105 CFU/ml were detected. Urine samples were analyzed for main uropathogens / groups of uropathogens using quantitative multiplex real-time PCR. Diagnostic characteristics of PCR were computed relative to the results of urine culture. Results. In total, 896 urine samples were tested. Of them, significant bacteriuria was found in 28 cases (3%). The frequency of detection of Escherichia coli was 50%, Enterococcus spp. — 25%, Klebsiella spp. — 7%, Proteus spp. and S. saprophyticus 4% each, Streptococcus spp. — 14%. Sensitivity and specificity of the detection of significant bacteriuria using quantitative real-time PCR for the majority of bacterial species / groups were 99% to 100%. Sensitivity and specificity of the quantitative real-time PCR based method were 96% and 98%, respectively. Conclusions. Prevalence of significant bacteriuria among pregnant women is 3%. Half of the uropathogens isolated from pregnant women with bacteriuria are E. coli. Sensitivity and specificity of quantitative PCR for the detection of significant bacteriuria are 96% and 98%, respectively.

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Development of an in-house real-time pcr to monitor the prevalence of macrolide-resistant Mycoplasma pneumoniae in Hong Kong

Pathology ◽

10.1016/j.pathol.2016.12.341 ◽

2017 ◽

Vol 49 ◽

pp. S116

Author(s):

Ho-Yin Lam ◽

Thomas Hin-Ching Wong ◽

Anthony Tsz-Chun Wong ◽

Gilman Kit-Hang Siu ◽

Wing-Cheong Yam ◽

...

Keyword(s):

Hong Kong ◽

Real Time ◽

Mycoplasma Pneumoniae ◽

Real Time Pcr

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Evaluation of the New BD Max GC Real-Time PCR Assay, Analytically and Clinically as a Supplementary Test for the BD ProbeTec GC Qx Amplified DNA Assay, for Molecular Detection of Neisseria gonorrhoeae

Journal of Clinical Microbiology ◽

10.1128/jcm.01962-15 ◽

2015 ◽

Vol 53 (12) ◽

pp. 3935-3937 ◽

Cited By ~ 5

Author(s):

Daniel Golparian ◽

Stina Boräng ◽

Martin Sundqvist ◽

Magnus Unemo

Keyword(s):

Neisseria Gonorrhoeae ◽

Real Time ◽

Sensitivity And Specificity ◽

Real Time Pcr ◽

Molecular Detection ◽

Analytical Sensitivity ◽

Pcr Assay ◽

Content Type ◽

Dna Assay ◽

Supplementary Test

The new BD Max GC real-time PCR assay showed high clinical and analytical sensitivity and specificity. It can be an effective and accurate supplementary test for the BD ProbeTec GC Qx amplified DNA assay, which had suboptimal specificity, and might also be used for initial detection ofNeisseria gonorrhoeae.

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