Rapid and Easy Identification of Capsular Serotypes of Streptococcus pneumoniae by Use of Fragment Analysis by Automated Fluorescence-Based Capillary Electrophoresis

Selva, Laura; Amo, Eva del; Brotons, Pedro; Muñoz-Almagro, Carmen

doi:10.1128/jcm.01368-12

Cited by 50 publications

(45 citation statements)

References 42 publications

(49 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A high throughput method for the identification of pneumococcal capsular types was established using multiplex PCR-CE (FAF-mPCR) [29]. The automation of the process can genotype 30 isolates in a few hours and carry out 90 reactions in 90 min, three-fold less time than that of Quellung reaction, these results indicated that FAFmPCR was a useful and alternative method to determine the capsular serotype of Streptococcus pneumoniae.…”

Section: Detecting and Diagnosing Bacteriamentioning

confidence: 88%

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Lian

Zhao

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

View full text Add to dashboard Cite

Rapid transmission, high morbidity, and mortality are the features of human infectious diseases caused by microorganisms, such as bacteria, fungi, and viruses. These diseases may lead within a short period of time to great personal and property losses, especially in regions where sanitation is poor. Thus, rapid diagnoses are vital for the prevention and therapeutic intervention of human infectious diseases. Several conventional methods are often used to diagnose infectious diseases, e.g. methods based on cultures or morphology, or biochemical tests based on metabonomics. Although traditional methods are considered gold standards and are used most frequently, they are laborious, time consuming, and tedious and cannot meet the demand for rapid diagnoses. Disease diagnosis using capillary electrophoresis methods has the advantages of high efficiency, high throughput, and high speed, and coupled with the different nucleic acid detection strategies overcomes the drawbacks of traditional identification methods, precluding many types of false positive and negative results. Therefore, this review focuses on the application of capillary electrophoresis based on nucleic detection to the diagnosis of human infectious diseases, and offers an introduction to the limitations, advantages, and future developments of this approach.

show abstract

Section: Detecting and Diagnosing Bacteriamentioning

confidence: 88%

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Lian

Zhao

2016

Clinical Chemistry and Laboratory Medicine (CCLM)

View full text Add to dashboard Cite

show abstract

“…DNA detection of pneumolysin ( ply ) and LytA gene by Real-Time PCR ( polymerase chain reaction ) in CSF was performed according to previously reported assays [21–24]. …”

Section: Methodsmentioning

confidence: 99%

Mannose-binding lectin-deficient genotypes as a risk factor of pneumococcal meningitis in infants

et al. 2017

Self Cite

View full text Add to dashboard Cite

ObjectivesThe objective of this study was to evaluate to evaluate the role of mannose-binding-lectin deficient genotypes in pneumococcal meningitis (PM) in children.MethodsWe performed a 16-year retrospective study (January 2001 to March 2016) including patients ≤ 18 years with PM. Variables including attack rate of pneumococcal serotype (high or low invasive capacity) and MBL2 genotypes associated with low serum MBL levels were recorded.ResultsForty-eight patients were included in the study. Median age was 18.5 months and 17/48 episodes (35.4%) occurred in children ≤ 12 months old. Serotypes with high-invasive disease potential were identified in 15/48 episodes (31.2%). MBL2 deficient genotypes accounted for 18.8% (9/48). Children ≤ 12 months old had a 7-fold risk (95% CI: 1.6–29.9; p < 0.01) of having a MBL2 deficient genotype in comparison to those > 12 months old. A sub-analysis of patients by age group revealed significant proportions of carriers of MBL2 deficient genotypes among those ≤ 12 months old with PM caused by opportunistic serotypes (54.5%), admitted to the PICU (Pediatric Intensive Care Unit) (46.7%) and of White ethnicity (35.7%). These proportions were significantly higher than in older children (all p<0.05).ConclusionsOur results suggest that differences in MBL2 genotype in children ≤12 months old affects susceptibility to PM, and it may have an important role in the episodes caused by non-high invasive disease potential serotypes.

show abstract

“…It is based on the use of 40 different primers pairs for the specific detection of 40 serotypes/serogroups and a pair of additional primers for detecting a fragment of the cpsA (wzg) gene, which is highly common to all capsular serotypes. So, this technique allows to detect all capsular types and to differentiate 40 serotypes/ serogroups [13]. On the other hand, in samples with a positive result between 30 and 35 cycles, serotypes were identified by a published multiplex real-time PCR methodology that detects the wzg gene and differentiates 21 serotypes [12].…”

Section: Serotype Identificationmentioning

confidence: 99%

Estimation of the invasive disease potential of Streptococcus pneumoniae in children by the use of direct capsular typing in clinical specimens

Amo

Selva

Sevilla

et al. 2014

Eur J Clin Microbiol Infect Dis

View full text Add to dashboard Cite

Traditionally, invasiveness indexes have been based on culture methods. We aimed to establish a new classification of the invasive disease potential of pneumococcal serotypes causing invasive pediatric disease in the era of conjugate vaccines in Catalonia, Spain, by adding capsular typing of Streptococcus pneumoniae in direct sample. Two samples of children attended at the University Hospital Sant Joan de Déu (Barcelona, Spain) between 2007 and 2011 were compared: a first sample of 358 children with invasive pneumococcal disease and a second sample of 402 pneumococcal nasopharyngeal carriers selected from 714 healthy children admitted for minor surgical procedures. The most common invasive serotypes were 1 (20.1 %, n = 72), 19A (13.9 %, n = 50), 3 (12.3 %, n = 44), and 7FA (7.5 %, n = 27), whereas the most common serotypes in carriage were 19A (8.7 %, n = 38), 10FC33C (7.8 %, n = 34), 6C (6.9 %, n = 30), and 19FBC (5.5 %, n = 24). We detected a rate of cocolonization of 26.4 % (n = 89) among the 336 samples serotyped in the carriers population. Serotypes 1, 3, and 7FA were significantly associated with high invasiveness. Serotypes 6C, 10FC33C, 23A, 35B, 19FBC, 21, 11AD, 15BC, 23B, 34, and 6A were significantly associated with low invasiveness. Our results proved that the use of molecular techniques in direct sample for both the detection and the capsular identification of Streptococcus pneumoniae is very useful to obtain a more accurate calculation of the invasiveness of the different pneumococcal serotypes.

show abstract

Rapid and Easy Identification of Capsular Serotypes of Streptococcus pneumoniae by Use of Fragment Analysis by Automated Fluorescence-Based Capillary Electrophoresis

Cited by 50 publications

References 42 publications

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Capillary electrophoresis based on nucleic acid detection for diagnosing human infectious disease

Mannose-binding lectin-deficient genotypes as a risk factor of pneumococcal meningitis in infants

Estimation of the invasive disease potential of Streptococcus pneumoniae in children by the use of direct capsular typing in clinical specimens

Contact Info

Product

Resources

About