Polymerase Chain Reaction: An  Important Tool for Early Diagnosis of  Leptospirosis Cases

Mullan, Summaiya; Panwala, Tanvi

doi:10.7860/jcdr/2016/22462.9010

Cited by 17 publications

(14 citation statements)

References 20 publications

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“…DNA of suggestive cultures with turbidity equivalent to 0.5 in the McFarland scale (1.5 x 10 8 bacteria/mL) was extracted through silica gel column with DNeasy Blood & Tissue ® kit (Qiagen ® , Germany), following the manufacturer instructions. The DNA was kept frozen at -20 °C until use.…”

Section: Methodsmentioning

confidence: 99%

Characterization of Leptospira isolates from humans and the environment in Uruguay

Meny

Menéndez

Quintero

et al. 2017

Rev. Inst. Med. trop. S. Paulo

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Laboratory diagnosis of human leptospirosis usually relies on indirect methods exploring specific immune response. Isolation and identification of the involved strains are cumbersome, but can provide biological resources for pathogenic studies and relevant information for guiding prevention and control measures. The aim of the research we are hereby reporting was the characterization of Leptospira isolates obtained from humans and the environment in Uruguay. Blood cultures were performed from early samples of 302 Uruguayan patients, mainly rural workers, and from 36 water samples taken from their living or working environments. Eight human isolates and seven environmental isolates were obtained and analyzed by end point Polymerase Chain Reaction (PCR), Multilocus Variable Number of Tandem Repeat Analysis (MLVA) and other molecular methods. Human isolates corresponded to several serogroups and serovars of Leptospira interrogans and Leptospira kirschneri species, probably reflecting the infection with similar involved Leptospira species and serovars of an extended animal reservoir in rural settings of the country, mostly dedicated to meat and dairy production. Culture-positive patients were older than usually affected workers, and presented signs and symptoms of severe illness. A high organic and circulating bacterial burden may explain an easier positive result from these workers’ samples. Environmental isolates were mainly identified as Leptospira biflexa strains, with a single L. meyeri isolate of uncertain significance.

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Section: Methodsmentioning

confidence: 99%

Characterization of Leptospira isolates from humans and the environment in Uruguay

Meny

Menéndez

Quintero

et al. 2017

Rev. Inst. Med. trop. S. Paulo

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“…The positivity of SAT and ELISA IgM for all 67 sera was 48 (71.6%) and 39 (58.2%), respectively. In the case of the unavailability of the reference serology test (MAT), ELISA IgM and SAT can detect leptospirosis at the acute and convalescent stages, with high sensitivity as reported (2,4,19,23,24). Therefore, we suggest that patients who had negative results by ELISA IgM and SAT might have not leptospirosis.…”

Section: Discussionmentioning

confidence: 77%

“…De Abreu Fonseca et al compared PCR with ELISA IgM, and reported that ELISA IgM had more sensitivity; however, PCR was most sensitive in initial sera samples presenting no specific antibodies detectable by any of the serological methods tested (18). Therefore, it was suggested that the positivity of the diagnosis increased when PCR was used with serological tests (18,23).…”

Section: Discussionmentioning

confidence: 99%

First Real-Time PCR in Morocco for Human Leptospirosis Using TaqMan Probes Targeting the LipL32 Gene

Al-orry

Arahou

Hassikou

et al. 2019

Arch Clin Infect Dis

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Background: Currently, qPCR has been used as a rapid diagnostic method for human leptospirosis. Previous studies have indicated that qPCR has high sensitivity in the early days of the illness. Objectives: The aim of this study was to evaluate qPCR as a diagnostic method for human leptospirosis in the National Institute of Hygiene, Rabat, Morocco. Methods: From 2004 to 2016, 67 sera related to 67 patients with clinical signs mimic to leptospirosis were sent to the laboratory of Bacteriology for routine diagnosis and confirmation. SAT, ELISA IgM, ELISA IgG, and qPCR were used for the diagnosis. Results: High positivity was observed by SAT (88.24%), ELISA IgM (58.82%), and real-time PCR (17.64%), in sequence. No negative results by serological tests had positive results by real-time PCR. Forty-six patients were males (68.68%) and 21 were females (31.34%). The high incidence observed was from Sidi Qacem (40%). Conclusions: SAT and ELISA IgM are useful for the diagnosis of human leptospirosis in Morocco and they can provide prompt and low-cost diagnosis, especially when resources are limited.

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“…Targeting housekeeping genes that are included in all species of Leptospira (16S rrs , secY , or gyrB ) or pathogenic species-specific genes ( lipL32 , lfb1 , or lig ) have been used as targets for PCR-based diagnosis. 2 10 11 12 13 The sensitivity and specificity of PCR has been reported as 52.7% and 97.2%, respectively, compared to MAT (49.8% and 98.8%, respectively). 3 PCR for the detection of pathogenic Leptospira spp.…”

Section: Discussionmentioning

confidence: 99%

Usefulness of Nested Polymerase Chain Reaction with Clinical Specimens for the Diagnosis of Leptospirosis: a Case Series and a Review of the Literature

Kim

et al. 2020

J Korean Med Sci

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A culture of the Leptospira species and the microscopic agglutination test (MAT) are considered as the reference standard for the diagnosis of leptospirosis, but both tests are imperfect for early diagnosis. We describe 4 patients diagnosed with leptospirosis using nested polymerase chain reaction (N-PCR) that targeted the 16S rRNA gene and the passive hemagglutination assay (PHA). In our 4 cases, Leptospira DNA in the urine, plasma, or cerebrospinal fluid (CSF), was detected by N-PCR in the early phase of leptospirosis, except in the sample from the buffy coat. Especially, case 3 showed that N-PCR with the urine and CSF was positive 8 days after symptom onset, but not for the plasma or buffy coat. We report 4 cases of leptospirosis that were diagnosed by N-PCR that targeted the 16S rRNA gene with urine, plasma, or CSF, but not the buffy coat. Three were cured by doxycycline but the case 4 was fatal. Detection of Leptospira DNA by PCR from the urine and CSF, in addition to plasma, may be helpful to confirm the diagnosis.

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Polymerase Chain Reaction: An Important Tool for Early Diagnosis of Leptospirosis Cases

Cited by 17 publications

References 20 publications

Characterization of Leptospira isolates from humans and the environment in Uruguay

Characterization of Leptospira isolates from humans and the environment in Uruguay

First Real-Time PCR in Morocco for Human Leptospirosis Using TaqMan Probes Targeting the LipL32 Gene

Usefulness of Nested Polymerase Chain Reaction with Clinical Specimens for the Diagnosis of Leptospirosis: a Case Series and a Review of the Literature

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