Highly Sensitive Loop-Mediated Isothermal Amplification for the Detection of <i>Leptospira</i>

Chen, Hua-Wei; Weissenberger, Giulia; Atkins, Erin; Chao, Chien‐Chung; Suputtamongkol, Yupin; Ching, Wei‐Mei

doi:10.1155/2015/147173

Cited by 29 publications

(25 citation statements)

References 25 publications

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“…Primer sets for the LAMP assay include the forward outer primer (F3), the backward outer primer (B3), the forward inner primer (FIP), the backward inner primer (BIP), and two other primers designed to accelerate the amplification, including the forward loop primer (LF) and the backward loop primer (LB). With the primers in hand, the LAMP assay can be performed with simple and readily available incubation sources like a water bath or heating block for isothermal heating [35,54,55].…”

Section: Principles Of Lamp Assaymentioning

confidence: 99%

“…Viruses 2019, 11, x FOR PEER REVIEW 2 of 20 backward loop primer (LB). With the primers in hand, the LAMP assay can be performed with simple and readily available incubation sources like a water bath or heating block for isothermal heating [35,54,55]. During the initial stages of the LAMP reaction, the inner primers (FIP or BIP) anneal to regions F2c or B2c within the target region.…”

Section: Principles Of Lamp Assaymentioning

confidence: 99%

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Loop-Mediated Isothermal Amplification (LAMP) for the Diagnosis of Zika Virus: A Review

Silva

Pardee

Pena

2019

Viruses

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The recent outbreak of Zika virus (ZIKV) in the Americas and its devastating developmental and neurological manifestations has prompted the development of field-based diagnostics that are rapid, reliable, handheld, specific, sensitive, and inexpensive. The gold standard molecular method for lab-based diagnosis of ZIKV, from either patient samples or insect vectors, is reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The method, however, is costly and requires lab-based equipment and expertise, which severely limits its use as a point-of-care (POC) tool in resource-poor settings. Moreover, given the lack of antivirals or approved vaccines for ZIKV infection, a POC diagnostic test is urgently needed for the early detection of new outbreaks and to adequately manage patients. Loop-mediated isothermal amplification (LAMP) is a compelling alternative to RT-qPCR for ZIKV and other arboviruses. This low-cost molecular system can be freeze-dried for distribution and exhibits high specificity, sensitivity, and efficiency. A growing body of evidence suggests that LAMP assays can provide greater accessibility to much-needed diagnostics for ZIKV infections, especially in developing countries where the ZIKV is now endemic. This review summarizes the different LAMP methods that have been developed for the virus and summarizes their features, advantages, and limitations.Viruses 2020, 12, 19 2 of 20 raising concerns about the neurological tropism of the virus [12]. In May 2015, the first case of ZIKV infection was reported in Brazil [13] and the virus rapidly spread throughout the country and much of Latin America, causing the largest recorded epidemic of the virus to date [14]. The Brazilian epidemic raised great international concern because of severe birth defects, including microcephaly, in neonates born to mothers infected by ZIKV during pregnancy [3,15,16].ZIKV is transmitted mainly through the bite of infected mosquitoes from the genus Aedes, although other vectors may also be involved in the transmission [17][18][19][20][21]. Additionally, other routes of ZIKV transmission have been identified, including blood transfusions, transplacental, perinatal, and sexual intercourse [22][23][24]. ZIKV infection usually causes a self-limiting and a mild illness, where the majority of cases are asymptomatic and, when present, symptoms include fever, headache, rash, conjunctivitis, and arthralgia [25]. In regions where there is a circulation of other arboviruses, such as DENV and chikungunya (CHIKV), the clinical diagnosis of ZIKV infection becomes extremely difficult because of common symptoms. Therefore, laboratory-based molecular diagnosis is of fundamental importance to correctly identify the etiologic agent [3,26].Given the lack of approved vaccines and antivirals against ZIKV, a rapid and reliable point-of-care (POC) diagnostic test for detection of ZIKV is urgently required for control and prevention measures and to increase the diagnostic capacity of ZIKV-affected, mainly in low-resource areas [27,28]. Z...

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Section: Principles Of Lamp Assaymentioning

confidence: 99%

Section: Principles Of Lamp Assaymentioning

confidence: 99%

Loop-Mediated Isothermal Amplification (LAMP) for the Diagnosis of Zika Virus: A Review

Silva

Pardee

Pena

2019

Viruses

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“…Loop-mediated isothermal amplification (LAMP) assays for Leptospira have been of interest for this purpose, as these do not require real-time PCR instruments [38]. Chen et al (2015) and Nurul Najian et al (2016) reported the design and analytical evaluation of two new LAMP assays for pathogenic Leptospira [32,33]. Both assays target lipL32 , while the assay by Chen et al includes lipL41 as a second target.…”

Section: Molecular Diagnostics For Leptospirosismentioning

confidence: 99%

“…Both assays target lipL32 , while the assay by Chen et al includes lipL41 as a second target. Both tests demonstrated good analytical performance; however neither publication included a clinical evaluation [32,33]. …”

Section: Molecular Diagnostics For Leptospirosismentioning

confidence: 99%

Molecular diagnostics for human leptospirosis

Waggoner

Pinsky²

2016

Current Opinion in Infectious Diseases

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Purpose of Review The definitive diagnosis of leptospirosis, which results from infection with spirochetes of the genus Leptospira, currently relies on the use of culture, serological testing (microscopic agglutination testing, MAT), and molecular detection. The purpose of this review is to describe new molecular diagnostics for Leptospira and discuss advancements in the use of available methods. Recent Findings Efforts have been focused on improving the clinical sensitivity of Leptospira detection using molecular methods. In this review, we describe a re-optimized pathogenic species-specific real-time PCR (targeting lipL32) that has demonstrated improved sensitivity; findings by two groups that real-time RT-PCRs targeting the 16S rrs gene can improve detection; and two new loop-mediated amplification techniques. Quantitation of leptospiremia, detection in different specimen types, and the complementary roles played by molecular detection and MAT will be discussed. Finally, a protocol for Leptospira strain subtyping using variable number tandem repeat targets and high-resolution melting will be described. Summary Molecular diagnostics have an established role for the diagnosis of leptospirosis and provide an actionable diagnosis in the acute setting. The use of real-time RT-PCR for testing serum/plasma and cerebrospinal fluid, when available, may improve the detection of Leptospira without decreasing clinical specificity.

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“…A combination of primer sets targeting lipL32 and lipL41 genes for Leptospira spp. detection of human specimens, shows positive results with PCR Leptospira –positive samples which are classified into three species including L. interrogans , L. borgpetersenii and L. weilii [ 5 ]. The study of Koizumi et al .…”

mentioning

confidence: 99%

Evaluation of loop-mediated isothermal amplification method (LAMP) for pathogenic <i>Leptospira</i> spp. detection with leptospires isolation and real-time PCR

Suwancharoen

Sittiwicheanwong

Wiratsudakul

2016

The Journal of Veterinary Medical Science

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Leptospirosis has been one of the worldwide zoonotic diseases caused by pathogenic Leptospira spp. Many molecular techniques have consecutively been developed to detect such pathogen including loop–mediated isothermal amplification method (LAMP). The objectives of this study were to evaluate the diagnostic accuracy of LAMP assay and real-time PCR using bacterial culture as the gold standard and to assess the agreement among these three tests using Cohen’s kappa statistics. In total, 533 urine samples were collected from 266 beef and 267 dairy cattle reared in central region of Thailand. Sensitivity and specificity of LAMP were 96.8% (95% CI 81.5–99.8) and 97.0% (95% CI 94.9–98.2), respectively. The accuracy of LAMP (97.0%) was significantly higher than that of real-time PCR (91.9%) at 95% CI. With Cohen’s kappa statistics, culture method and LAMP were substantially agreed with each other (77.4%), whereas real-time PCR only moderately agreed with culture (47.7%) and LAMP (45.3%), respectively. Consequently, LAMP was more effective than real-time PCR in detecting Leptospira spp. in the urine of cattle. Besides, LAMP had less cost and was simpler than real-time PCR. Thus, LAMP was an excellent alternative for routine surveillance of leptospirosis in cattle.

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Highly Sensitive Loop-Mediated Isothermal Amplification for the Detection of Leptospira

Cited by 29 publications

References 25 publications

Loop-Mediated Isothermal Amplification (LAMP) for the Diagnosis of Zika Virus: A Review

Loop-Mediated Isothermal Amplification (LAMP) for the Diagnosis of Zika Virus: A Review

Molecular diagnostics for human leptospirosis

Evaluation of loop-mediated isothermal amplification method (LAMP) for pathogenic <i>Leptospira</i> spp. detection with leptospires isolation and real-time PCR

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