The search for improved methods for diagnosing leptospirosis: the approach of a laboratory in Brescia, Italy

Ml, Pacciarini; Ml, Savio; Donini, G.; Tagliabue, Silvia

doi:10.20506/rst.12.2.711

Cited by 2 publications

(2 citation statements)

References 42 publications

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“…Several primer pairs for PCR detection of leptospires have been described, some based on specific gene targets (483), most frequently 16S or 23S rRNA genes (287,386,407,607,637,639,667) and repetitive elements (428,502,643,670,671), while others have been constructed from genomic libraries (247,248,324,594). However, few have been shown to amplify leptospiral DNA from either human (247,386) or veterinary (378,559,594,606,643,670) clinical material, and of these, only two methods have been subject to extensive clinical evaluation (84,387).…”

Section: Molecular Diagnosismentioning

confidence: 99%

Leptospirosis

2001

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Leptospirosis is a worldwide zoonotic infection with a much greater incidence in tropical regions and has now been identified as one of the emerging infectious diseases. The epidemiology of leptospirosis has been modified by changes in animal husbandry, climate, and human behavior. Resurgent interest in leptospirosis has resulted from large outbreaks that have received significant publicity. The development of simpler, rapid assays for diagnosis has been based largely on the recognition that early initiation of antibiotic therapy is important in acute disease but also on the need for assays which can be used more widely. In this review, the complex taxonomy of leptospires, previously based on serology and recently modified by a genotypic classification, is discussed, and the clinical and epidemiological value of molecular diagnosis and typing is also evaluated

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Section: Molecular Diagnosismentioning

confidence: 99%

Leptospirosis

2001

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“…Targets comprised the genes lig [51], lipL32/hap1 [52], lipL21 and lipL32 [53,54] and lipL41 [55]. Other approaches comprised the increase of the sensitivity of the PCR by using primer pairs targeting repetitive elements [56][57][58][59][60], or of both sensitivity and specificity by applying nested PCR primers [61,62] or using a subsequent southern hybridization step with a specific internal [32, 39,42,44]. Whereas a multitude of conventional PCRs have been described, only few have been subjected to clinical evaluation at a limited scale.…”

Section: Molecular Detection: Diagnosismentioning

confidence: 99%

Molecular Approaches in the Detection and Characterization of Leptospira

Ahmed¹,

Grobusch²

2012

J Bacteriol Parasitol

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Conventional diagnosis of leptospirosis and characterization of Leptospira spp. relies mainly on serology. A major drawback of serology in diagnosis is that it needs sufficient levels of anti-Leptospira antibodies, thus jeopardizing confirmation in the early acute phase of disease. The cross agglutinin absorption test (CAAT) that determines Leptospira serovars is a technically demanding and laborious method and therefore is only performed at a few laboratories. Novel molecular diagnostic tests mainly rely on the polymerase chain reaction (PCR). PCR perfectly complements serological testing, since it is especially sensitive in the first 5 days after onset of symptoms. Real time PCR is rapid and has been validated for their high clinical accuracy. The introduction of molecular techniques for defining Leptospira species revolutionized the categorization of strains in this genus, as species and serogroups appeared to show little correlation. The reference test in molecular speciation is based on determining DNA homology. This approach is tedious and user-unfriendly and therefore is increasingly replaced by other techniques. To date, a wealth of molecular typing methods is available. Most attractive are those techniques that provide direct digital and electronically portable data. Such techniques comprise fluorescent amplified fragment length polymorphism (FAFLP), array typing, multilocus variable number of tandem repeats analysis (MLVA) and sequence-based phylogeny, and to some extent pulsed field gel electrophoresis (PFGE). Multilocus sequence typing (MLST) is the most robust method for determining Leptospira strain diversity and in the future will probably only be surpassed by phylogeny on whole genome sequences.

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The search for improved methods for diagnosing leptospirosis: the approach of a laboratory in Brescia, Italy

Cited by 2 publications

References 42 publications

Leptospirosis

Leptospirosis

Molecular Approaches in the Detection and Characterization of Leptospira

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