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rapid detection of mycoplasma pneumoniae by loop-mediated isothermal amplification (lamp) in clinical respiratory specimens
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نویسنده
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arfaatabar maryam ,noori goodarzi narjes ,afshar davoud ,memariani hamed ,azimi ghasem ,masoorian ensieh ,pourmand mohammad reza
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منبع
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iranian journal of public health - 2019 - دوره : 48 - شماره : 5 - صفحه:917 -924
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چکیده
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Background: mycoplasma pneumoniae is a common cause of community-acquired pneumonia (cap) worldwide, especially among children and debilitated populations. the present study aimed to investigate a loop-mediated isothermal amplification (lamp) technique for rapid detection of m. pneumoniae in clinical specimens collected from patients with pneumonia. methods: throat swabs were collected from 110 outpatients who suffered from pneumonia. throat swab samples were obtained from patients referred to the hospital outpatient clinics of tehran university hospitals, iran in 2017. the presence of m. pneumoniae in the clinical specimens was evaluated by lamp, pcr and culture methods. sensitivity and specificity of the lamp and pcr assays were also determined. results: out of 110 specimens, lamp assay detected m. pneumoniae in 35 specimens. detection limit of the lamp assay was determined to be 33fg /μl or ~ 40 genome copies/reaction. moreover, no cross-reaction with genomic dna from other bacteria was observed. only 25 specimens were positive by the culture method. the congruence between lamp assay and culture method was ‘substantial’ (κ=0.77). specificity and sensitivity of lamp assay were 88.2%, 100% in compare with culture. however, the congruence between lamp assay and pcr assay was ‘almost perfect’ (κ=0.86). specificity and sensitivity of lamp assay were 92.5%, 100% in compare with pcr. conclusion: overall, the lamp assay is a rapid and cost-efficient laboratory test in comparison to other methods including pcr and culture. therefore, the lamp method can be applied in identification of m. pneumoniae isolates in respiratory specimens.
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کلیدواژه
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mycoplasma pneumoniae ,pcr ,loop-mediated isothermal amplification (lamp) ,culture
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آدرس
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tehran university of medical sciences, school of public health, department of pathobiology, ایران, tehran university of medical sciences, school of public health, department of pathobiology, ایران, zanjan university of medical sciences, school of medicine, department of microbiology and virology, ایران, pasteur institute of iran, biotechnology research center, ایران, shahed university of medical sciences, department of internal medicine, ایران, tehran university of medical sciences, school of public health, department of pathobiology, ایران, tehran university of medical sciences, school of public health, biotechnology research center, department of pathobiology, ایران
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پست الکترونیکی
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mpourmand@tums.ac.ir
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Authors
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