Development of a new multiplex PCR to detect prevalent species of house dust mites in house dust

Oliveira, Ana Sofia; Gaspar, Carlos; Rolo, Joana; Pereira, Cristiana; Palmeira‐de‐Oliveira, Rita; Teixeira, João Paulo; Martinez-de-Oliveira, José; Palmeira‐de‐Oliveira, Ana

doi:10.1080/09603123.2021.1893282

Cited by 2 publications

(2 citation statements)

References 31 publications

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“…Oliveira et al ( 2021 ) designed an rDNA-based mPCR capable of distinguishing D. pteronyssinus and D. farinae . The method was tested on five dust samples and, to assess specificity, they sequenced only two mites, one per species.…”

Section: Discussionmentioning

confidence: 99%

“…Due to these drawbacks, molecular techniques based on ribosomal DNA (rDNA) and cytochrome oxydase subunit 1 ( coxI ) have been used as important tools on mite identification and phylogenetic analysis (Lava Kumar et al 1999 ; Navajas and Fenton 2000 ; Vargas et al 2005 ; Ben-David et al 2007 ; De Rojas et al 2007 ; Osakabe et al 2008 ; Wong et al 2011 ; Yang et al 2011 ; Matsuda et al 2012 ; Thet-Em et al 2012 ; Arimoto et al 2013 ; Roussel et al 2013 ; Stephenson et al 2013 ; Navia et al 2014 ; Beroiz et al 2014 ; Khaing et al 2014 ; Lv et al 2014 ; Juan et al 2015 ; Li et al 2015 ; Stalažs and Moročko-Bičevska 2016 ; Peltier et al 2018 ; Syromyatnikov et al 2018 ; Zélé et al 2018 ; Zhou et al 2020 ; Gómez-Martínez et al 2020 ; Oliveira et al 2021 ; Zhang et al 2021 ).…”

Section: Introductionmentioning

confidence: 99%

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Identification of Glycycometus malaysiensis (for the first time in Brazil), Blomia tropicalis and Dermatophagoides pteronyssinus through multiplex PCR

et al. 2022

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Blomia tropicalis and Dermatophagoides pteronyssinus play an important role in triggering allergy . Glycycometus malaysiensis causes IgE reaction in sensitive people, but is rarely reported in domestic dust, because it is morphologically similar to B. tropicalis making the identification of these species difficult. The identification of mites is mostly based on morphology, a time-consuming and ambiguous approach. Herein, we describe a multiplex polymerase chain reaction (mPCR) assay based on ribosomal DNA capable to identify mixed cultures of B. tropicalis , D. pteronyssinus and G. malaysiensis , and/or to identify these species from environmental dust. For this, the internal transcribed spacer 2 (ITS2) regions, flanked by partial sequences of the 5.8S and 28S genes, were PCR-amplified, cloned and sequenced. The sequences obtained were aligned with co-specific sequences available in the GenBank database for primer design and phylogenetic studies. Three pairs of primers were chosen to compose the mPCR assay, which was used to verify the frequency of different mites in house dust samples ( n = 20) from homes of Salvador, Brazil. Blomia tropicalis was the most frequent, found in 95% of the samples, followed by G. malaysiensis (70%) and D. pteronyssinus (60%). Besides reporting for the first time the occurrence of G. malaysiensis in Brazil, our results confirm the good resolution of the ITS2 region for mite identification. Furthermore, the mPCR assay proved to be a fast and reliable tool for identifying these mites in mixed cultures and could be applied in future epidemiological studies, and for quality control of mite extract production for general use. Supplementary Information The online version contains supplementary material available at 10.1007/s10493-022-00694-y.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Identification of Glycycometus malaysiensis (for the first time in Brazil), Blomia tropicalis and Dermatophagoides pteronyssinus through multiplex PCR

et al. 2022

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Multiplex LAMP assay for detecting the prevalent species of dust mites Dermatophagoides farinae and Dermatophagoides pteronyssinus in the domestic environment

Shuai,

Xue,

et al. 2024

Sci Rep

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Dermatophagoides farina (D. farinae) and Dermatophagoides pteronyssinus (D. pteronyssinus) are the prevalent kinds of house dust mites (HDMs). HDMs are common inhalant allergens that cause a range of allergic diseases, such as rhinitis, atopic dermatitis, and asthma. The epidemiology of these diseases is associated with exposure to mites. Therefore, in the present study, a method named multiplex loop-mediated isothermal amplification (LAMP) was developed to detect environmental dust mites. The multiplex LAMP assay allows amplification within a single tube and has an ITS plasmid detection limit as low as 40 fg/µL for both single dust mites and mixed dust mites (D. pteronyssinus and D. farinae), which is up to ten times more sensitive than classical PCR techniques. Furthermore, the multiplex LAMP method was applied to samples of single dust mites and clinical dust to confirm its validity. The multiplex LAMP assay exhibited higher sensitivity, simpler instrumentation, and visualization of test results, indicating that this method could be used as an alternative to traditional techniques for the detection of HDMs.

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Development of a new multiplex PCR to detect prevalent species of house dust mites in house dust

Cited by 2 publications

References 31 publications

Identification of Glycycometus malaysiensis (for the first time in Brazil), Blomia tropicalis and Dermatophagoides pteronyssinus through multiplex PCR

Identification of Glycycometus malaysiensis (for the first time in Brazil), Blomia tropicalis and Dermatophagoides pteronyssinus through multiplex PCR

Multiplex LAMP assay for detecting the prevalent species of dust mites Dermatophagoides farinae and Dermatophagoides pteronyssinus in the domestic environment

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