Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid and Accurate Confirmatory Diagnosis of HTLV-1/2 Infection

Gomes, Yago Côrtes Pinheiro; Caterino–de–Araujo, Adele; Campos, Karoline Rodrigues; Gonçalves, Maria Gisele; Leite, Ana Cláudia Celestino Bezerra; Lima, Marco Antônio; Araújo, Abelardo; Silva, Marcus Tulius T.; Espíndola, Otávio de Melo

doi:10.3390/v12090981

Cited by 18 publications

(16 citation statements)

References 66 publications

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“…The finding of indeterminate results in the WB for HTLV and positive by PCR has already been reported in other studies (Broutet et al, 1996;Osti et al, 1998;Ji et al, 2020;Halbrook et al, 2021). About this phenomenon, one hypothesis that we can consider is a longer time for seroconversion that can be associated with low antigenic stimulation due to the defective proviruses (Cánepa et al, 2015;Kuramitsu et al, 2017;Gomes et al, 2020). Therefore, this sample was considered HTLV positive due to confirmation by nested-PCR.…”

Section: Discussionsupporting

confidence: 71%

Human T-Lymphotropic Virus-1/2 Infection in Central Brazil Prisons: A Multicenter Study

et al. 2022

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Human T-lymphotropic virus (HTLV) infection is endemic in some group populations. Prisoners are at high risk of acquiring HTLV infection mainly due to the environment of closed penal institutions, socioeconomic conditions, and risk behaviors. This study aimed to investigate the prevalence, the occurrence of new cases of HTLV infection after a 1-year follow-up, and circulating subtypes of HTLV-1 among prisoners from twelve closed prisons in Mato Grosso do Sul state, Central Brazil. A total of 1,507 prisoners were randomly enrolled in the cross-sectional study. Out of the 1,507 participants, 1,000 prisoners, susceptible to HTLV infection, were included in the prospective cohort study. In the cross-sectional study, serological evidence of HTLV infection was 0.4% (CI 95%: 0.1-0.7), tested for anti-HTLV antibodies by enzyme-linked immunosorbent assay (ELISA). Three samples were positive for HTLV-1, two samples were positive for HTLV-2, and 1 sample was indeterminate by the Western blot method. The presence of HTLV proviral DNA was detected in all positive samples by amplification of the HTLV tax gene through nested-PCR. Phylogenetic analysis showed that HTLV-1 samples belonged to the Cosmopolitan subtype Transcontinental subgroup. From a cohort of 1,000 individuals, no new case of HTLV infection was detected. Although the prevalence rate of HTLV infection found in this study was similar to that observed in the Brazilian general population, the lack of access to preventive interventions and harm reduction measures all contribute to increasing the risk of HTLV transmission and acquisition among this key and vulnerable population.

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Section: Discussionsupporting

confidence: 71%

Human T-Lymphotropic Virus-1/2 Infection in Central Brazil Prisons: A Multicenter Study

et al. 2022

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“…LAMP, as a low-cost, fast, simple, and efficient nucleic acid amplification technique, seems to be more satisfactory. Presently, LAMP- and LAMP-based assay have been applied in the detection of various pathogens (containing viruses, bacteria, and fungi) ( Sharma et al, 2016 ; Gomes et al, 2020 ). However, conventional validation tools for LAMP amplicons, including visual indicators, agarose gel electrophoresis, and real-time turbidimeter, are difficult to achieve simple, rapid, and accurate detection of target pathogens ( Li et al, 2019a ; Yang et al, 2021 ).…”

Section: Discussionmentioning

confidence: 99%

A Label-Based Polymer Nanoparticles Biosensor Combined with Loop-Mediated Isothermal Amplification for Rapid, Sensitive, and Highly Specific Identification of Brucella abortus

Yang

Wang

Liu

et al. 2021

Front. Bioeng. Biotechnol.

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Brucella abortus (B. abortus), an important zoonotic pathogen in Brucella spp., is the major causative agent of abortion in cattle (namely, bovine brucellosis). Currently, although the isolation and identification of the Brucella abortus were commonly accepted as the gold standard method, it cannot meet the requirements for early diagnostic strategies. Conventional PCR techniques and immunological tests can realize rapid detection of B. abortus, but the demands for PCR thermal cyclers and/or specific antibodies hinder their application in basic laboratories. Thus, rapid, sensitive, and specific diagnostic strategies are essential to prevent and control the spread of the bovine brucellosis. In this work, a novel detection method for the rapid identification of B. abortus, which uses loop-mediated isothermal amplification (LAMP) combined with a label-based polymer nanoparticles lateral flow immunoassay biosensor (LFIA), was established. One set of specific B. abortus-LAMP primers targeting the BruAb2_0168 gene was designed by the online LAMP primer design tool. The B. abortus-LAMP-LFIA assay was optimized and evaluated using various pathogens and whole blood samples. The optimal amplification temperature and time for B. abortus-LAMP-LFIA were determined to be 65°C and 50 min, respectively. The B. abortus-LAMP-LFIA method limit of detection (LoD) was 100 fg per reaction for pure genomic DNA of B. abortus. Meanwhile, the detection specificity was 100%, and there was no cross-reactivity for other Brucella members and non-Brucella strains. Furthermore, the entire procedure, including the DNA preparation for whole blood samples (30 min), isothermal incubation (50 min), and LFIA detection (2–5 min), can be completed in approximately 85 min. Thus, the B. abortus-LAMP-LFIA assay developed was a simple, rapid, sensitive, and reliable detection technique, which can be used as a screening and/or diagnostic tool for B. abortus in the field and basic laboratories.

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“…The specific detection of pathogenic bacteria is of great significance for disease prevention and control [ 45 , 48 , 49 ]. In this study, 37 types of strains were used to evaluate the accuracy and sensitivity of primers.…”

Section: Discussionmentioning

confidence: 99%

First Report of Crown Gall of Kiwifruit (Actinidia deliciosa) Caused by Agrobacterium fabacearum in China and the Establishment of Loop-Mediated Isothermal Amplification Technique

Shi

Zhang

et al. 2021

IJMS

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Kiwifruit is moderately sweet and sour and quite popular among consumers; it has been widely planted in some areas of the world. In 2019, the crown gall disease of kiwifruit was discovered in the main kiwifruit-producing area of Guizhou Province, China. This disease can weaken and eventually cause the death of the tree. The phylogeny, morphological and biological characteristics of the bacteria were described, and were related to diseases. The pathogenicity of this species follows the Koch hypothesis, confirming that A. fabacearum is the pathogen of crown gall disease of kiwifruit in China. In this study, Loop-mediated isothermal amplification (LAMP) analysis for genome-specific gene sequences was developed for the specific detection of A. fabacearum. The detection limit of the LAMP method is 5 × 10−7 ng/μL, which has high sensitivity. At the same time, the amplified product is stained with SYBR Green I after the reaction is completed, so that the amplification can be detected with the naked eye. LAMP analysis detected the presence of A. fabacearum in the roots and soil samples of the infected kiwifruit plant. The proposed LAMP detection technology in this study offers the advantages of ease of operation, visibility of results, rapidity, accuracy and high sensitivity, making it suitable for the early diagnosis of crown gall disease of kiwifruit.

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Loop-Mediated Isothermal Amplification (LAMP) Assay for Rapid and Accurate Confirmatory Diagnosis of HTLV-1/2 Infection

Cited by 18 publications

References 66 publications

Human T-Lymphotropic Virus-1/2 Infection in Central Brazil Prisons: A Multicenter Study

Human T-Lymphotropic Virus-1/2 Infection in Central Brazil Prisons: A Multicenter Study

A Label-Based Polymer Nanoparticles Biosensor Combined with Loop-Mediated Isothermal Amplification for Rapid, Sensitive, and Highly Specific Identification of Brucella abortus

First Report of Crown Gall of Kiwifruit (Actinidia deliciosa) Caused by Agrobacterium fabacearum in China and the Establishment of Loop-Mediated Isothermal Amplification Technique

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