Microbiological comparison of blood culture and amplification of 16S rDNA methods in combination with DGGE for detection of neonatal sepsis in blood samples

García-Gudiño, Isela; Yllescas-Medrano, Eucario; Maida-Claros, Rolando; Soriano-Becerril, Diana; Díaz, Néstor Fabián; García‐López, Guadalupe; Molina‐Hernández, Anayansí; Flores-Herrera, Óscar; Serna, Francisco Javier Zavala-Díaz de la; Peralta-Pérez, María del Rosario; Flores‐Herrera, Héctor

doi:10.1007/s00431-017-3036-3

Cited by 5 publications

(6 citation statements)

References 37 publications

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“…We applied the QUADAS-2 tool to evaluate the methodological quality of included studies against microbiological culture reference standard (see Figure 2 ). For a few studies in the patient selection domain ( 34 , 38 , 40 , 48 , 53 , 55 , 63 ), the risk of bias was high because the studies could not prevent improper sample exclusion. Regarding the applicability concern, all studies included blood samples from patients suspected of BSI, indicating a low risk of bias.…”

Section: Resultsmentioning

confidence: 99%

“…Eight studies ( 21 , 32 , 38 , 48 , 51 , 55 , 57 , 59 ) met the inclusion criteria for comparing NAAT with a microbiological culture among children consisting of a total of 4,089 samples for MSSA detection in suspected BSI patients. The sensitivity and specificity of NAAT for MSSA detection ranged from 0.25 (95% CI 0.0–0.94) to 1.0 (95% CI, 0.85–1.0), whilst our search did not result in any MRSA studies ( Figure 3 ).…”

Section: Resultsmentioning

confidence: 99%

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Clinical Utility of Molecular Tests for Guiding Therapeutic Decisions in Bloodstream Staphylococcal Infections: A Meta-Analysis

et al. 2021

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Background: Treatment of bloodstream staphylococcal infections (BSI) necessitates the prompt initiation of appropriate antimicrobial agents and the rapid de-escalation of excessive broad-spectrum coverage to reduce the risk of mortality. We, therefore, aimed to demonstrate the diagnostic accuracy of nucleic acid amplification tests (NAAT) for the identification of methicillin-resistant S. aureus (MRSA) and methicillin-sensitive S. aureus (MSSA) in clinically suspected patients.Methods: Until November 23, 2020, databases including PubMed, Scopus, Embase, and Web of Science were scanned for eligible studies. A bivariate random-effects model was used for meta-analysis of the 33 included studies obtained from 1606 citations, and pooled summary estimates with 95% confidence intervals (CI) were generated.Results: Twenty-three studies (n = 8,547) assessed NAAT accuracy for MSSA detection, while three studies (n = 479) evaluated MRSA detection in adults. The pooled NAAT sensitivity and specificity for MRSA in adults was higher [sensitivity: 0.83 (95% CI 0.59–0.96), specificity: 0.99 (95% CI 0.98–1.0)] as compared to MSSA [sensitivity: 0.76 (95% CI 0.69–0.82), specificity: 0.98 (95% CI 0.98–0.99)]. Similarly, eight studies (n = 4,089) investigating MSSA in pediatric population reported higher NAAT accuracy [sensitivity: 0.89 (95% CI 0.76–0.96), specificity: 0.98 (95% CI 0.97–0.98)] compared to adults. Among NAA tests, SeptiFast (real-time PCR, commercial) was frequently applied, and its diagnostic accuracy corresponded well to the overall summary estimates. A meta-regression and subgroup analysis of study design, sample condition, and patient selection method could not explain the heterogeneity (P > 0.05) in the diagnostic efficiency.Conclusions: NAAT could be applied as the preferred initial tests for timely diagnosis and BSI management.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Clinical Utility of Molecular Tests for Guiding Therapeutic Decisions in Bloodstream Staphylococcal Infections: A Meta-Analysis

et al. 2021

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“…Bone marrow and blood samples have been generally considered to be aseptic body fluids. The positive amplification would be of great reference value for infectious diseases [11]. The ITS in the fungal ribose is conservative, which has been widely used in the identification of the fungal species and the phylogenetic researches [13].…”

Section: Discussionmentioning

confidence: 99%

“…PCR was performed with the Leishmania- [9], Histoplasma capsulatum- [10] and Penicillium marneffei-specific [10] primers, respectively, and no specific bands were observed. Moreover, the 16S rDNA V3 [11] amplification showed negative results.…”

Section: Case Reportmentioning

confidence: 99%

Molecular diagnosis and source tracing of an infection of Aureobasidium pullulans

Liu¹,

Shi

et al. 2019

J Infect Dev Ctries

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Aureobasidium pullulans (A. pullulans) is a dematiaceous, yeast-like fungus that is ubiquitous in nature, which can colonize the human hair and skin. A. pullulans has been clinically implicated to cause skin and soft tissue infections, meningitis, splenic abscesses, and peritonitis. Herein, molecular diagnostic of internal transcribed spacer (ITS) sequencing was used to investigate a suspected case of A. pullulans infection, and the infection source had been traced. A 27-year-old female case was suspected of kala-azar due to the recurrent fever. Bone marrow specimens were analyzed. The samples were negative for Leishmania, Penicillium marneffei and Histoplasma capsulatum. DNA was extracted from the bone marrow specimens, and the 583-bp sequence was amplified with the fungal ITS universal primers. The sequence was compared by Blast query to be identified as A. pullulans. A strain of A. pullulans was also isolated from the kitchen of the patient’s living room. Culture characteristics were the same as the human pathogens of A. pullulans, and the ITS sequence was identical to the bone marrow ITS amplification. In conclusion, a deep infection caused by A. pullulans is rare, often occurring in the indwelling catheter, which may cause peritonitis and other symptoms. ITS sequencing of fungi can be used as a diagnostic reference. As A. pullulans is a common fungus in environment, amplification of ITS sequence of A. pullulans in the aseptic body fluid would be necessary to make a comprehensive diagnosis based on the clinical symptoms and signs.

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“…In developed countries, the estimated prevalence is 2-8% (24). The Department of Neonatal Intensive Care of the National Institute of Perinatology "Isidro Espinosa de los Reyes" (INPerIER) in Mexico City has reported an incidence of 2.3% in the total number of births attended during a 5-year period (26).…”

Section: Introductionmentioning

confidence: 99%

Increased Levels of Plasma Extracellular Heat-Shock Proteins 60 and 70 kDa Characterized Early-Onset Neonatal Sepsis

et al. 2021

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Background: Extracellular heat-shock proteins (eHsp) are highly conserved molecules that play an important role in inflammatory diseases and have been quantified in plasma from patients with infectious diseases, including sepsis. There is a constant search for dependable biochemical markers that, in combination with conventional methods, could deliver a prompt and reliable diagnosis of early-onset neonatal sepsis.Objective: We sought to assess the level of eHsp-27, eHsp-60, eHsp-70, and tumor necrosis factor-alpha (TNFα) in plasma of healthy neonates at term and infants with early-onset neonatal sepsis.Methods: This study included 34 newborns that were classified as healthy neonates at term (blood samples from the umbilical cord, n = 23) or infants with early-onset neonatal sepsis (blood samples obtained from umbilical artery by standard sterile procedures before starting a systemic antibiotic intervention, n = 11). All blood samples were centrifuged, and the plasma recovered to determine eHsp-27, eHsp-60, eHsp-70, and TNFα levels by ELISA.Results: Our results indicate that the level of eHsp-27 in healthy neonates at term was 0.045 ± 0.024 pg/ml. This value decreased 2.5-fold in infants with early-onset neonate sepsis (0.019 ± 0.006 pg/ml, p = 0.004). In contrast, the levels of eHsp-60 and eHsp-70 in healthy neonates at term were 13.69 ± 5.3 and 4.03 ± 2.6 pg/ml, respectively. These protein levels increased significantly 1.8- and 1.9-fold in the plasma of infants with early-onset neonatal sepsis (p ≤ 0.001). The level of TNFα in healthy neonates at term was 2.94 ± 0.46 pg/ml, with a 3.0-fold increase in infants with early-onset neonatal sepsis (8.96 ± 0.72 pm/ml, p ≤ 0.001). The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of eHsp compared with that of C-reactive protein were 73.3, 60.0, 47.8, and 33.3%, respectively.Conclusion: This study demonstrated a consistent increase of eHsp-60 and eHsp-70 in the plasma of infants diagnosed with early-onset neonatal sepsis. These proteins showed higher sensitivity and specificity than C-reactive protein and blood culture test.

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Microbiological comparison of blood culture and amplification of 16S rDNA methods in combination with DGGE for detection of neonatal sepsis in blood samples

Cited by 5 publications

References 37 publications

Clinical Utility of Molecular Tests for Guiding Therapeutic Decisions in Bloodstream Staphylococcal Infections: A Meta-Analysis

Clinical Utility of Molecular Tests for Guiding Therapeutic Decisions in Bloodstream Staphylococcal Infections: A Meta-Analysis

Molecular diagnosis and source tracing of an infection of Aureobasidium pullulans

Increased Levels of Plasma Extracellular Heat-Shock Proteins 60 and 70 kDa Characterized Early-Onset Neonatal Sepsis

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