dgkh000509
10.3205/dgkh000509
urn:nbn:de:0183-dgkh0005093
Research Article
An analysis of the efficacy of universal PCR and BACTEC 9120 BD for identifying bacteremia in pediatrics
Ein Vergleich der Wirksamkeit von universeller PCR und BACTEC 9120 BD zur Identifizierung einer Bakteriämie in der Pädiatrie
Safarkhani
Safarkhani
Azam
A
Pediatric Infections Research Center, Research Institute for Children’s Health, Shahid Beheshti University of Medical Sciences, Tehran, Iran
author
Shirkavand
Shirkavand
Fatemeh
F
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
author
Abdollahi
Abdollahi
Nafiseh
N
Pediatric Infections Research Center, Research Institute for Children’s Health, Shahid Beheshti University of Medical Sciences, Tehran, Iran
author
Ahari Oskooie
Ahari Oskooie
Nazanin
N
Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
author
Azimi
Azimi
Leila
L
Dr.
Pediatric Infections Research Center, Research Institute of Children’s Health, Shahid Beheshti, University of Medical Sciences, Tehran, Iran, P.O. Box 15468-15514, Iran, Phone: +98 21 22227004Pediatric Infections Research Center, Research Institute for Children’s Health, Shahid Beheshti University of Medical Sciences, Tehran, Iran
leilaazimi1982@gmail.com
author
Rahbar
Rahbar
Mohammad
M
Department of Microbiology, Reference Health Laboratories Research Center, Ministry of Health and Medical Education, Tehran, Iran
author
Karimi
Karimi
Abdollah
A
Pediatric Infections Research Center, Research Institute for Children’s Health, Shahid Beheshti University of Medical Sciences, Tehran, Iran
author
German Medical Science GMS Publishing House
Düsseldorf
610
bacteremia
BACTEC
molecular methods
phenotypic methods
Bakterämie
BACTEC
molekulare Methoden
phenotypische Methoden
20241023
engl
This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 License.
Dieser Artikel ist ein Open-Access-Artikel und steht unter den Lizenzbedingungen der Creative Commons Attribution 4.0 License (Namensnennung).
2196-5226
19
GMS Hygiene and Infection Control
GMS Hyg Infect Control
54
Shahid Behehsti University of Medical Sciences
Hintergrund: Blutstrominfektionen (BSI) gehört zu den schwerwiegenden Erkrankungen in der Pädiatrie und können die Morbidität und Mortalität erhöhen. Blutkulturen sind zeitaufwändig und können in manchen Fällen falsch-negative Ergebnisse liefern, wie z.B. bei intrazellulären oder anspruchsvollen Bakterien. Ziel dieser Studie war es, die PCR im Vergleich zur automatisierten Blutkultur mit BACTEC zu bewerten.Materialien und Methode: In der Querschnittsstudie wurden Blutproben von hospitalisierten Kindern im Mofid-Kinderkrankenhaus mit Anzeichen einer Bakteriämie von Februar bis Mai 2023 erfasst. Die Bakterien wurden mittels phänotypischer und PCR-Methode identifiziert.Ergebnisse: Es wurden 150 Blutproben untersucht. 60% bzw. 40% der Proben wiesen bei beiden Methoden negative bzw. positive Ergebnisse auf. Die PCR zeigte eine 100%ige Sensitivität und Spezifit<![CDATA[ät beim Nac</PlainText></TextGroup>hweis einer Bakteriämie im Vergleich zu BACTEC. Eine Vielzahl von Bakterien wurde durch phänotypische und molekulare Methoden identifiziert, wobei Koagulase-negative <Mark2>Staphylococcus</Mark2> (CoNS) den größten Anteil ausmachten.</Pgraph><Pgraph><Mark1>Schlussfolgerung:</Mark1> Die wichtigsten Vorteile des molekularen Tests sind der rasche und genaue Nachweis von bakteriellen Krankheitserregern mit hoher Sensitivität und Spezifität. </Pgraph></Abstract>
<Abstract language="en" linked="yes"><Pgraph><Mark1>Background:</Mark1> Bloodstream infections (BSI) are serious diseases in pediatrics and can increase the rate of morbidity and mortality. Blood culture is time consuming and can have false negative results in some case such as the intracellular or fastidious bacteria. This study aimed to evaluate the PCR against automated blood culture with BACTEC.</Pgraph><Pgraph><Mark1>Materials and methods:</Mark1> In this observational cross-sectional study the blood samples of hospitalized children in Mofid Children’s Hospital with bacteremia signs from February to May 2023 were enrolled. The causative bacteria in bacteremia were identified by phenotypic and PCR methods. </Pgraph><Pgraph><Mark1>Results:</Mark1> 150 blood samples were enrolled to identify the presence of bacteremia by BACTEC and PCR. 60% and 40% of samples have negative and positive results in both methods, respectively. PCR showed 100% sensitivity and specificity in detecting bacteremia compared to BACTEC. A variety of bacteria were identified by phenotypic and molecul<TextGroup><PlainText>ar me</PlainText></TextGroup>thods and coagulase negative <Mark2>Staphylococcus</Mark2> (CONS) is the most of them.</Pgraph><Pgraph><Mark1>Conclusion:</Mark1> The rapid and accurate detection of bacterial pathogens with the high sensitivity and specificity compared gold standard method are the most important profits of molecular assay.</Pgraph></Abstract>
<TextBlock linked="yes" name="Introduction">
<MainHeadline>Introduction</MainHeadline><Pgraph>Nosocomial infections (NI) are related with different toxins or infectious agents such as bacteria, viruses or parasites that cause infection among patients admitt<TextGroup><PlainText>ed to the h</PlainText></TextGroup>ospital <TextLink reference="1"></TextLink>. The most common form of NI is blood infection or bacteremia <TextLink reference="2"></TextLink> that increases in-hospital mortality and increases treatment costs, especially in ill newborns who are admitted to the NICU or ill children that in PICU <TextLink reference="3"></TextLink>, <TextLink reference="4"></TextLink>. Patients with hemato-oncologic diseases are at high risk for developing bacteremia because they are often severely immunocompromised due to underlying disease, antineoplastic therapy, and/or hematopoietic stem cell transplantation. This infection associated with some kind of invasive device entering the venous blood system such as central vascular catheters (CVC) and lead to the central line-associated bloodstream infection <TextLink reference="5"></TextLink>, <TextLink reference="6"></TextLink>. Bacteremia is now associated with significant morbidity and mortality in pediatrics and the appropriate and prompt treatment of this infection is necessary and has been shown to significantly reduce mortality <TextLink reference="7"></TextLink>. In order to accurate<TextGroup><PlainText>ly id</PlainText></TextGroup>entify the microorganisms responsible for these infections, blood culture stands out as a crucial diagnostic procedure. It is recommended that blood samples be collected just before initiating any empirical antimicrobial therapy. Nonetheless, the effectiveness of blood culture may be compromised in cases where patients have recently been on antibiotics or when dealing with slow-growing or intracellular micro-organisms, leading to delays and lower sensitivity in detection. Identifying microorganisms, particularly in blood samples, through the utilization of pathogen-specific or broad-spectrum PCR tests shows great potential. Furthermore, the advancements in Real-Time PCR technology come with a multitude of benefits when compared to conventional PCR methods. These advantages encompass rapid results, ease of use, accurate quantification abilities, and reduced chances of contamination <TextLink reference="8"></TextLink>. The bloodstream infection (BSI) is an important infection in children, especially in hospitalized patients in PICU and can increase the rate of morbidity and mortality <TextLink reference="9"></TextLink>, <TextLink reference="10"></TextLink>. BSI specially affects the outcome of children undergoing cardiac surgery, increasing complications and mortality <TextLink reference="9"></TextLink>. For this reason, it is important to improve BSI diagnosis. So, the purpose of this research is to analyze results of PCR against of BACTEC in samples sent to the Children’s Infectious Research Center (PIRC) Laboratory at Mofid Children's Hospital.</Pgraph></TextBlock>
<TextBlock linked="yes" name="Materials and methods">
<MainHeadline>Materials and methods</MainHeadline><SubHeadline>Sample preparation and set up</SubHeadline><Pgraph>The research was conducted as an observational cross-sectional study at Mofid Children’s Hospital from February to May 2023. The study included collecting blood samples from hospitalized patients in the Pediatric Intensive Care Unit (PICU), Neonatal Intensive Care Unit (NICU), as well as patients in the transplant and hematology units showing symptoms of bacteremia like Fever and chills. The study was approved by the ethics committees of the research institute for children health, Shahid Behehshti University of Medical Science by approved ID: <TextGroup><PlainText>IR.SBMU.RICH.1402.009</PlainText></TextGroup>. </Pgraph><SubHeadline>BACTEC process</SubHeadline><Pgraph>BACTEC bottles (BD BACTEC™ 9120) were sent from wards prefilled with 5 ml of whole blood to laboratory of PIRC. The bottles were incubated immediately upon receipt in the microbiology laboratory in accordance with the manufacturer’s recommendation. Bottles flagged as positive by the BACTEC system were sub-cultured in MacConkey and blood agar Following that identified by gram stain and other biochemical tests like oxidase, catalase, TSI, and DNase.</Pgraph><SubHeadline>Molecular detection</SubHeadline><Pgraph>300 µl of each BACTEC samples were used for extracting total DNA by commercial extraction kit (SIMBIOLAB). The extracted samples were used for conventional PCR for detecting 16S rRNA in all samples for confirmation of present of bacteria in samples. In the next step for detecting the <Mark2>Staphylococcus aureus, Enterococcus spp., </Mark2><TextGroup><Mark2>Esch</Mark2></TextGroup><Mark2>erichia coli, Klebsiella pneumoniae</Mark2> and <Mark2>Enterobacter cloacae</Mark2> conventional PCR as well as Real Time PCR was used for detecting <Mark2>Neisseria meningitidis, Hemophilus influenzae</Mark2> and <Mark2>Streptococcus pneumoniae</Mark2>. The primers were shown on Table 1 <ImgLink imgNo="1" imgType="table"/>. </Pgraph><SubHeadline>Statistical analysis</SubHeadline><Pgraph>The data was analyzed using statistical software SPSS version 27. P value of less than 0.05 was accepted <TextGroup><PlainText>at the</PlainText></TextGroup> level of significance.</Pgraph></TextBlock>
<TextBlock linked="yes" name="Results">
<MainHeadline>Results</MainHeadline><Pgraph>150 blood samples from hospitalized children under <TextGroup><PlainText>18 years</PlainText></TextGroup> old were enrolled to identify the presence of bacteremia by both BACTEC and PCR methods. 90 (60%) out of 150 samples had negative results in both BACTEC and PCR assay as well as 60 (40%) samples were positive. This result indicates a strong correlation between results of PCR and BACTEC, with PCR demonstrating 100% sensitivity and specificity in detecting bacteremia compared to bacteria culture as a gold standard technique.</Pgraph><Pgraph>The bacteria were identified by phenotypic identification following a positive BACTEC. Molecular analysis was then conducted on samples that tested positive for 16srRNA in PCR. The most identified bacteria in both methods was coagulase negative <Mark2>Staphylococcus</Mark2> (CONS) (Table 2 <ImgLink imgNo="2" imgType="table"/>). </Pgraph><Pgraph>The phenotypic approach was unable to identify any of co-infection, but PCR analysis confirmed the presence of co-infections with <Mark2>K. pneumonia</Mark2> and <Mark2>E. cloacae</Mark2> 2 cases.</Pgraph><Pgraph>Two co-infections with <Mark2>Klebsiella pneumoniae</Mark2> and <Mark2>Enterobacter cloacae</Mark2> were detected by PCR. While the <TextGroup><Mark2>Citrob</Mark2></TextGroup><Mark2>acter</Mark2> spp. and <Mark2>Klebsiella pneumoniae</Mark2> were identified in phenotypic methods in these two samples. </Pgraph><Pgraph>In one instance, <Mark2>Staphylococcus aureus</Mark2> was identified using a phenotypics methods but PCR detected the CONS. </Pgraph><Pgraph>A study reported the presence of <Mark2>Pseudomonas</Mark2> spp. and <Mark2>Acinetobacter</Mark2> spp. using a phenotypic method, but the genus of the species was not clearly defined. However, through PCR analysis, it was possible to identify <Mark2>P. aeruginosa</Mark2> and <Mark2>A. baumannii</Mark2>. Specifically, 11 samples were identified as <Mark2>P. aeruginosa</Mark2>, 4 as non-<Mark2>aerugino</Mark2><TextGroup><Mark2>sa Pseu</Mark2></TextGroup><Mark2>domonas</Mark2>, 8 as <Mark2>A. baumannii</Mark2>, and 3 as non-<Mark2>baumannii Acinetobacter</Mark2> using PCR analysis.</Pgraph></TextBlock>
<TextBlock linked="yes" name="Discussion">
<MainHeadline>Discussion</MainHeadline><Pgraph>Due to the low sensitivity of blood cultures, it is cruci<TextGroup><PlainText>al t</PlainText></TextGroup>o develop new approaches to quickly identify the bacteria responsible for infections, especially in children <TextLink reference="9"></TextLink>. Traditional techniques like phenotypic tests can be replaced with innovative methods such as multiple PCR because the phenotypic methods are time-consuming and occasionally have false negative results <TextLink reference="9"></TextLink>. Molecular technologies enable quick and precise identification of the pathogen responsible for infectious diseases in a much shorter duration compared to conventional methods <TextLink reference="11"></TextLink>, <TextLink reference="12"></TextLink>.</Pgraph><Pgraph>The findings of this study have validated the superi<TextGroup><PlainText>or ac</PlainText></TextGroup>curacy of PCR as a molecular diagnostic tool for detecting bacteremia. PCR can amplify various regions of the bacterial 16S ribosomal ribonucleic acid (rRNA) and accurately identify the causative bacteria. Our study demonstrated that PCR conducted directly on whole blood samples correlates well with BACTEC, consistent with findings from previous studies <TextLink reference="13"></TextLink>, <TextLink reference="14"></TextLink>. A study has highlighted a crucial need for the ability to access PCR test results for antimicrobial stewardship purposes within hospital settings <TextLink reference="15"></TextLink>, <TextLink reference="16"></TextLink>. The findings from these studies <TextLink reference="15"></TextLink>, <TextLink reference="16"></TextLink> align with our research results. </Pgraph><Pgraph>The latest findings from this study reveal that there was no discernible distinction in detecting Gram-positive and Gram-negative bacteria responsible for caus<TextGroup><PlainText>ing b</PlainText></TextGroup>acteremia. All samples exhibited a strong correlation with the outcomes obtained from the BACTEC bacterial culture technique.</Pgraph><Pgraph>PCR is more effective at identifying bacteria responsible for bacteremia than BACTEC. Sometimes, determining the genus of bacteria requires complex testing procedures and the use of resources therefore, the laboratory can identify specific strains of bacteria by PCR. For instance, the laboratory mentioned in this research study can only identify <Mark2>Pseudomonas</Mark2> spp. or <Mark2>Acinetobacter</Mark2> spp. However, by using PCR, we can easily detect the genus of bacteria. Another crucial aspect is the use of molecular assays for accurately identifying bacteria in co-infections. Our findings indicate that PCR successfully identified two co-infections in bloodstream infections, phenotyp<TextGroup><PlainText>ic iden</PlainText></TextGroup>tification methods did not. The key advantages of using molecular testing for bacteremia diagnosis include rapid and accurate detection of causative bacteria, making PCR a viable alternative to bacterial culture and phenotypic identification methods.</Pgraph></TextBlock>
<TextBlock linked="yes" name="Conclusion">
<MainHeadline>Conclusion</MainHeadline><Pgraph>As technology continues to advance, molecular techniques are playing a more prominent role in standard microbiological testing and have brought about a significant change in how we deal with bloodstream infections. Being able to quickly diagnose severe infections is crucial for starting the right treatment promptly and avoiding the unnecessary use of antibiotics, which can lead to adverse effects and increased medical expenses. The key benefits of utilizing molecular methods for identifying bacterial pathogens and antimicrobial resistance genes include faster and more cost-effective results.</Pgraph></TextBlock>
<TextBlock linked="yes" name="Notes">
<MainHeadline>Notes</MainHeadline><SubHeadline>Authors’ ORCID </SubHeadline><Pgraph><UnorderedList><ListItem level="1">Azam Safarkhani: <Hyperlink href="https://orcid.org/0009-0003-3325-498X">0009-0003-3325-498X</Hyperlink></ListItem><ListItem level="1">Fatemeh Shirkavand: <Hyperlink href="https://orcid.org/0009-0001-5207-4883">0009-0001-5207-4883</Hyperlink></ListItem><ListItem level="1">Nafiseh Abdollahi: <Hyperlink href="https://orcid.org/0000-0002-1820-4680">0000-0002-1820-4680</Hyperlink></ListItem><ListItem level="1">Nazanin Ahari Oskooie: <Hyperlink href="https://orcid.org/0009-0008-5647-7525">0009-0008-5647-7525</Hyperlink></ListItem><ListItem level="1">Leila Azimi: <Hyperlink href="https://orcid.org/0000-0002-7216-2530">0000-0002-7216-2530</Hyperlink></ListItem><ListItem level="1">Mohammad Rahbar: <Hyperlink href="https://orcid.org/0000-0002-3070-3108">0000-0002-3070-3108</Hyperlink></ListItem><ListItem level="1">Abdollah Karimi: <Hyperlink href="https://orcid.org/0000-0002-4225-0097">0000-0002-4225-0097</Hyperlink></ListItem></UnorderedList></Pgraph><SubHeadline>Ethical approval</SubHeadline><Pgraph>The ethical approval number of this study is (<TextGroup><PlainText>IR.SBMU.RICH.REC.1402.009</PlainText></TextGroup>) from the Ethics Committee of the Research Institute for Children’s Health, Shahid <TextGroup><PlainText>Beheshti</PlainText></TextGroup> University of Medical Sciences, Tehran, Iran. </Pgraph><SubHeadline>Data Availability Statement</SubHeadline><Pgraph>The data presented in this study are available upon reasonable request from the corresponding author.</Pgraph><SubHeadline>Funding</SubHeadline><Pgraph>Research reported in this publication was supported by Researcher Grant Committee under grant number [43006041] from the Shahid Behehsti Universi<TextGroup><PlainText>ty of Me</PlainText></TextGroup>dical Sciences, Tehran, Iran.</Pgraph><SubHeadline>Competing interests</SubHeadline><Pgraph>The authors declare that they have no competing interests.</Pgraph></TextBlock>
<References linked="yes">
<Reference refNo="1">
<RefAuthor>Nimer NA</RefAuthor>
<RefTitle>Nosocomial infection and antibiotic-resistant threat in the Middle East</RefTitle>
<RefYear>2022</RefYear>
<RefJournal>Infect Drug Resist</RefJournal>
<RefPage>631-9</RefPage>
<RefTotal>Nimer NA. Nosocomial infection and antibiotic-resistant threat in the Middle East. Infect Drug Resist. 2022 Feb 25;15:631-9. DOI: 10.2147/IDR.S351755</RefTotal>
<RefLink>https://doi.org/10.2147/IDR.S351755</RefLink>
</Reference>
<Reference refNo="2">
<RefAuthor>Jeong IS</RefAuthor>
<RefAuthor>Jeong JS</RefAuthor>
<RefAuthor>Choi EO</RefAuthor>
<RefTitle>Nosocomial infection in a newborn intensive care unit (NICU), South Korea</RefTitle>
<RefYear>2006</RefYear>
<RefJournal>BMC Infect Dis</RefJournal>
<RefPage>103</RefPage>
<RefTotal>Jeong IS, Jeong JS, Choi EO. Nosocomial infection in a newborn intensive care unit (NICU), South Korea. BMC Infect Dis. 2006 Jun 23;6:103. DOI: 10.1186/1471-2334-6-103</RefTotal>
<RefLink>https://doi.org/10.1186/1471-2334-6-103</RefLink>
</Reference>
<Reference refNo="3">
<RefAuthor>Hudome SM</RefAuthor>
<RefAuthor>Fisher MC</RefAuthor>
<RefTitle>Nosocomial infections in the neonatal intensive care unit</RefTitle>
<RefYear>2001</RefYear>
<RefJournal>Curr Opin Infect Dis</RefJournal>
<RefPage>303-7</RefPage>
<RefTotal>Hudome SM, Fisher MC. Nosocomial infections in the neonatal intensive care unit. Curr Opin Infect Dis. 2001 Jun;14:303-7. DOI: 10.1097/00001432-200106000-00010</RefTotal>
<RefLink>https://doi.org/10.1097/00001432-200106000-00010</RefLink>
</Reference>
<Reference refNo="4">
<RefAuthor>Kawagoe JY</RefAuthor>
<RefAuthor>Segre CA</RefAuthor>
<RefAuthor>Pereira CR</RefAuthor>
<RefAuthor>Cardoso MF</RefAuthor>
<RefAuthor>Silva CV</RefAuthor>
<RefAuthor>Fukushima JT</RefAuthor>
<RefTitle>Risk factors for nosocomial infections in critically ill newborns: a 5-year prospective cohort study</RefTitle>
<RefYear>2001</RefYear>
<RefJournal>Am J Infect Control</RefJournal>
<RefPage>109-14</RefPage>
<RefTotal>Kawagoe JY, Segre CA, Pereira CR, Cardoso MF, Silva CV, Fukushima JT. Risk factors for nosocomial infections in critically ill newborns: a 5-year prospective cohort study. Am J Infect Control. 2001 Apr;29:109-14. DOI: 10.1067/mic.2001.114162</RefTotal>
<RefLink>https://doi.org/10.1067/mic.2001.114162</RefLink>
</Reference>
<Reference refNo="5">
<RefAuthor>Sohn AH</RefAuthor>
<RefAuthor>Garrett DO</RefAuthor>
<RefAuthor>Sinkowitz-Cochran RL</RefAuthor>
<RefAuthor>Grohskopf LA</RefAuthor>
<RefAuthor>Levine GL</RefAuthor>
<RefAuthor>Stover BH</RefAuthor>
<RefAuthor>Siegel JD</RefAuthor>
<RefAuthor>Jarvis WR</RefAuthor>
<RefAuthor> Pediatric Prevention Network</RefAuthor>
<RefTitle>Prevalence of nosocomial infections in neonatal intensive care unit patients: Results from the first national point-prevalence survey</RefTitle>
<RefYear>2001</RefYear>
<RefJournal>J Pediatr</RefJournal>
<RefPage>821-7</RefPage>
<RefTotal>Sohn AH, Garrett DO, Sinkowitz-Cochran RL, Grohskopf LA, Levine GL, Stover BH, Siegel JD, Jarvis WR; Pediatric Prevention Network. Prevalence of nosocomial infections in neonatal intensive care unit patients: Results from the first national point-prevalence survey. J Pediatr. 2001 Dec;139(6):821-7. DOI: 10.1067/mpd.2001.119442</RefTotal>
<RefLink>https://doi.org/10.1067/mpd.2001.119442</RefLink>
</Reference>
<Reference refNo="6">
<RefAuthor>Baier C</RefAuthor>
<RefAuthor>Linke L</RefAuthor>
<RefAuthor>Eder M</RefAuthor>
<RefAuthor>Schwab F</RefAuthor>
<RefAuthor>Chaberny IF</RefAuthor>
<RefAuthor>Vonberg RP</RefAuthor>
<RefAuthor>Ebadi E</RefAuthor>
<RefTitle>Incidence, risk factors and healthcare costs of central line-associated nosocomial bloodstream infections in hematologic and oncologic patients</RefTitle>
<RefYear>2020</RefYear>
<RefJournal>PLoS One</RefJournal>
<RefPage>e0227772</RefPage>
<RefTotal>Baier C, Linke L, Eder M, Schwab F, Chaberny IF, Vonberg RP, Ebadi E. Incidence, risk factors and healthcare costs of central line-associated nosocomial bloodstream infections in hematologic and oncologic patients. PLoS One. 2020 Jan 24;15(1):e0227772. DOI: 10.1371/journal.pone.0227772</RefTotal>
<RefLink>https://doi.org/10.1371/journal.pone.0227772</RefLink>
</Reference>
<Reference refNo="7">
<RefAuthor>Irwin AD</RefAuthor>
<RefAuthor>Drew RJ</RefAuthor>
<RefAuthor>Marshall P</RefAuthor>
<RefAuthor>Nguyen K</RefAuthor>
<RefAuthor>Hoyle E</RefAuthor>
<RefAuthor>Macfarlane KA</RefAuthor>
<RefAuthor>Wong HF</RefAuthor>
<RefAuthor>Mekonnen E</RefAuthor>
<RefAuthor>Hicks M</RefAuthor>
<RefAuthor>Steele T</RefAuthor>
<RefAuthor>Gerrard C</RefAuthor>
<RefAuthor>Hardiman F</RefAuthor>
<RefAuthor>McNamara PS</RefAuthor>
<RefAuthor>Diggle PJ</RefAuthor>
<RefAuthor>Carrol ED</RefAuthor>
<RefTitle>Etiology of childhood bacteremia and timely antibiotics administration in the emergency department</RefTitle>
<RefYear>2015</RefYear>
<RefJournal>Pediatrics</RefJournal>
<RefPage>635-42</RefPage>
<RefTotal>Irwin AD, Drew RJ, Marshall P, Nguyen K, Hoyle E, Macfarlane KA, Wong HF, Mekonnen E, Hicks M, Steele T, Gerrard C, Hardiman F, McNamara PS, Diggle PJ, Carrol ED. Etiology of childhood bacteremia and timely antibiotics administration in the emergency department. Pediatrics. 2015 Apr;135(4):635-42. DOI: 10.1542/peds.2014-2061</RefTotal>
<RefLink>https://doi.org/10.1542/peds.2014-2061</RefLink>
</Reference>
<Reference refNo="8">
<RefAuthor>Fenollar F</RefAuthor>
<RefAuthor>Raoult D</RefAuthor>
<RefTitle>Molecular diagnosis of bloodstream infections caused by non-cultivable bacteria</RefTitle>
<RefYear>2007</RefYear>
<RefJournal>Int J Antimicrob Agents</RefJournal>
<RefPage>S7-15</RefPage>
<RefTotal>Fenollar F, Raoult D. Molecular diagnosis of bloodstream infections caused by non-cultivable bacteria. Int J Antimicrob Agents. 2007 Nov;30(Suppl 1):S7-15. DOI: 10.1016/j.ijantimicag.2007.06.024</RefTotal>
<RefLink>https://doi.org/10.1016/j.ijantimicag.2007.06.024</RefLink>
</Reference>
<Reference refNo="9">
<RefAuthor>Checa RMC</RefAuthor>
<RefAuthor>Gijón M</RefAuthor>
<RefAuthor>Hofheinz SB</RefAuthor>
<RefAuthor>Rojo P</RefAuthor>
<RefTitle>Multiple polymerase chain reaction for direct detection of bloodstream infection after cardiac surgery in a PICU</RefTitle>
<RefYear>2022</RefYear>
<RefJournal>Crit Care Explor</RefJournal>
<RefPage>e0707</RefPage>
<RefTotal>Checa RMC, Gijón M, Hofheinz SB, Rojo P. Multiple polymerase chain reaction for direct detection of bloodstream infection after cardiac surgery in a PICU. Crit Care Explor. 2022 May 27;4(6):e0707. DOI: 10.1097/CCE.0000000000000707</RefTotal>
<RefLink>https://doi.org/10.1097/CCE.0000000000000707</RefLink>
</Reference>
<Reference refNo="10">
<RefAuthor>Elward AM</RefAuthor>
<RefAuthor>Fraser VJ</RefAuthor>
<RefTitle>Risk factors for nosocomial primary bloodstream infection in pediatric intensive care unit patients: a 2-year prospective cohort study</RefTitle>
<RefYear>2006</RefYear>
<RefJournal>Infect Control Hosp Epidemiol</RefJournal>
<RefPage>553-60</RefPage>
<RefTotal>Elward AM, Fraser VJ. Risk factors for nosocomial primary bloodstream infection in pediatric intensive care unit patients: a 2-year prospective cohort study. Infect Control Hosp Epidemiol. 2006 Jun;27(6):553-60. DOI: 10.1086/505096</RefTotal>
<RefLink>https://doi.org/10.1086/505096</RefLink>
</Reference>
<Reference refNo="17">
<RefAuthor>Campos-Peña E</RefAuthor>
<RefAuthor>Martín-Nuñez E</RefAuthor>
<RefAuthor>Pulido-Reyes G</RefAuthor>
<RefAuthor>Martín-Padrón J</RefAuthor>
<RefAuthor>Caro-Carrillo E</RefAuthor>
<RefAuthor>Donate-Correa J</RefAuthor>
<RefAuthor>Lorenzo-Castrillejo I</RefAuthor>
<RefAuthor>Alcoba-Flórez J</RefAuthor>
<RefAuthor>Machín F</RefAuthor>
<RefAuthor>Méndez-Alvarez S</RefAuthor>
<RefTitle>Multiplex PCR assay for identification of six different Staphylococcus spp. and simultaneous detection of methicillin and mupirocin resistance</RefTitle>
<RefYear>2014</RefYear>
<RefJournal>J Clin Microbiol</RefJournal>
<RefPage>2698-701</RefPage>
<RefTotal>Campos-Peña E, Martín-Nuñez E, Pulido-Reyes G, Martín-Padrón J, Caro-Carrillo E, Donate-Correa J, Lorenzo-Castrillejo I, Alcoba-Flórez J, Machín F, Méndez-Alvarez S. Multiplex PCR assay for identification of six different Staphylococcus spp. and simultaneous detection of methicillin and mupirocin resistance. J Clin Microbiol. 2014 Jul;52(7):2698-701. DOI: 10.1128/JCM.00918-14</RefTotal>
<RefLink>https://doi.org/10.1128/JCM.00918-14</RefLink>
</Reference>
<Reference refNo="18">
<RefAuthor>Mwikuma G</RefAuthor>
<RefAuthor>Kainga H</RefAuthor>
<RefAuthor>Kallu SA</RefAuthor>
<RefAuthor>Nakajima C</RefAuthor>
<RefAuthor>Suzuki Y</RefAuthor>
<RefAuthor>Hang'ombe BM</RefAuthor>
<RefTitle>Determination of the Prevalence and Antimicrobial Resistance of Enterococcus faecalis and Enterococcus faecium Associated with Poultry in Four Districts in Zambia</RefTitle>
<RefYear>2023</RefYear>
<RefJournal>Antibiotics (Basel)</RefJournal>
<RefPage>657</RefPage>
<RefTotal>Mwikuma G, Kainga H, Kallu SA, Nakajima C, Suzuki Y, Hang'ombe BM. Determination of the Prevalence and Antimicrobial Resistance of Enterococcus faecalis and Enterococcus faecium Associated with Poultry in Four Districts in Zambia. Antibiotics (Basel). 2023 Mar 28;12(4):657. DOI: 10.3390/antibiotics12040657</RefTotal>
<RefLink>https://doi.org/10.3390/antibiotics12040657</RefLink>
</Reference>
<Reference refNo="19">
<RefAuthor>Gómez-Duarte OG</RefAuthor>
<RefAuthor>Arzuza O</RefAuthor>
<RefAuthor>Urbina D</RefAuthor>
<RefAuthor>Bai J</RefAuthor>
<RefAuthor>Guerra J</RefAuthor>
<RefAuthor>Montes O</RefAuthor>
<RefAuthor>Puello M</RefAuthor>
<RefAuthor>Mendoza K</RefAuthor>
<RefAuthor>Castro GY</RefAuthor>
<RefTitle>Detection of Escherichia coli enteropathogens by multiplex polymerase chain reaction from children's diarrheal stools in two Caribbean-Colombian cities</RefTitle>
<RefYear>2010</RefYear>
<RefJournal>Foodborne Pathog Dis</RefJournal>
<RefPage>199-206</RefPage>
<RefTotal>Gómez-Duarte OG, Arzuza O, Urbina D, Bai J, Guerra J, Montes O, Puello M, Mendoza K, Castro GY. Detection of Escherichia coli enteropathogens by multiplex polymerase chain reaction from children's diarrheal stools in two Caribbean-Colombian cities. Foodborne Pathog Dis. 2010 Feb;7(2):199-206. doi: 10.1089/fpd.2009.0355</RefTotal>
<RefLink>https://doi.org/10.1089/fpd.2009.0355</RefLink>
</Reference>
<Reference refNo="20">
<RefAuthor>Clarridge JE 3rd</RefAuthor>
<RefTitle>Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases</RefTitle>
<RefYear>2004</RefYear>
<RefJournal>Clin Microbiol Rev</RefJournal>
<RefPage>840-62, table of contents</RefPage>
<RefTotal>Clarridge JE 3rd. Impact of 16S rRNA gene sequence analysis for identification of bacteria on clinical microbiology and infectious diseases. Clin Microbiol Rev. 2004 Oct;17(4):840-62, table of contents. DOI: 10.1128/CMR.17.4.840-862.2004</RefTotal>
<RefLink>https://doi.org/10.1128/CMR.17.4.840-862.2004</RefLink>
</Reference>
<Reference refNo="21">
<RefAuthor>Zhejiang Institute of Freshwater Fisheries</RefAuthor>
<RefTitle></RefTitle>
<RefYear>2012</RefYear>
<RefBookTitle>Enterobacter cloacae specific PCR (polymerase chain reaction) detection primer</RefBookTitle>
<RefPage></RefPage>
<RefTotal>FiZhejiang Institute of Freshwater Fisheries. Enterobacter cloacae specific PCR (polymerase chain reaction) detection primer. 2012.</RefTotal>
</Reference>
<Reference refNo="22">
<RefAuthor>Karimi A</RefAuthor>
<RefAuthor>Rafiei Tabatabaei S</RefAuthor>
<RefAuthor>Azimi L</RefAuthor>
<RefAuthor>Almasian Tehrani N</RefAuthor>
<RefAuthor>Fallah F</RefAuthor>
<RefAuthor>Faghihian I</RefAuthor>
<RefTitle>Tracing the Negative Results of Multiplex Real-Time PCR Assay for Diagnosis of Bacterial Pediatrics Meningitis</RefTitle>
<RefYear>2023</RefYear>
<RefJournal>Can J Infect Dis Med Microbiol</RefJournal>
<RefPage>3502666</RefPage>
<RefTotal>Karimi A, Rafiei Tabatabaei S, Azimi L, Almasian Tehrani N, Fallah F, Faghihian I. Tracing the Negative Results of Multiplex Real-Time PCR Assay for Diagnosis of Bacterial Pediatrics Meningitis. Can J Infect Dis Med Microbiol. 2023 Jan 16;2023:3502666. DOI: 10.1155/2023/3502666</RefTotal>
<RefLink>https://doi.org/10.1155/2023/3502666</RefLink>
</Reference>
<Reference refNo="23">
<RefAuthor>Okafor JU</RefAuthor>
<RefAuthor>Nwodo UU</RefAuthor>
<RefTitle>Antibiogram Profile and Detection of Resistance Genes in Pseudomonas aeruginosa Recovered from Hospital Wastewater Effluent</RefTitle>
<RefYear>2023</RefYear>
<RefJournal>Antibiotics (Basel)</RefJournal>
<RefPage>1517</RefPage>
<RefTotal>Okafor JU, Nwodo UU. Antibiogram Profile and Detection of Resistance Genes in Pseudomonas aeruginosa Recovered from Hospital Wastewater Effluent. Antibiotics (Basel). 2023 Oct 6;12(10):1517. DOI: 10.3390/antibiotics12101517</RefTotal>
<RefLink>https://doi.org/10.3390/antibiotics12101517</RefLink>
</Reference>
<Reference refNo="24">
<RefAuthor>Xu L</RefAuthor>
<RefAuthor>Deng S</RefAuthor>
<RefAuthor>Wen W</RefAuthor>
<RefAuthor>Tang Y</RefAuthor>
<RefAuthor>Chen L</RefAuthor>
<RefAuthor>Li Y</RefAuthor>
<RefAuthor>Zhong G</RefAuthor>
<RefAuthor>Li J</RefAuthor>
<RefAuthor>Ting WJ</RefAuthor>
<RefAuthor>Fu B</RefAuthor>
<RefTitle>Molecular typing, and integron and associated gene cassette analyses in Acinetobacter baumannii strains isolated from clinical samples</RefTitle>
<RefYear>2020</RefYear>
<RefJournal>Exp Ther Med</RefJournal>
<RefPage>1943-52</RefPage>
<RefTotal>Xu L, Deng S, Wen W, Tang Y, Chen L, Li Y, Zhong G, Li J, Ting WJ, Fu B. Molecular typing, and integron and associated gene cassette analyses in Acinetobacter baumannii strains isolated from clinical samples. Exp Ther Med. 2020 Sep;20(3):1943-52. DOI: 10.3892/etm.2020.891</RefTotal>
<RefLink>https://doi.org/10.3892/etm.2020.891</RefLink>
</Reference>
<Reference refNo="11">
<RefAuthor>Gerace E</RefAuthor>
<RefAuthor>Mancuso G</RefAuthor>
<RefAuthor>Midiri A</RefAuthor>
<RefAuthor>Poidomani S</RefAuthor>
<RefAuthor>Zummo S</RefAuthor>
<RefAuthor>Biondo C</RefAuthor>
<RefTitle>Recent Advances in the Use of Molecular Methods for the Diagnosis of Bacterial Infections</RefTitle>
<RefYear>2022</RefYear>
<RefJournal>Pathogens</RefJournal>
<RefPage>663</RefPage>
<RefTotal>Gerace E, Mancuso G, Midiri A, Poidomani S, Zummo S, Biondo C. Recent Advances in the Use of Molecular Methods for the Diagnosis of Bacterial Infections. Pathogens. 2022 Jun 8;11(6):663. DOI: 10.3390/pathogens11060663</RefTotal>
<RefLink>https://doi.org/10.3390/pathogens11060663</RefLink>
</Reference>
<Reference refNo="12">
<RefAuthor>Tsalik EL</RefAuthor>
<RefAuthor>Bonomo RA</RefAuthor>
<RefAuthor>Fowler VG Jr</RefAuthor>
<RefTitle>New Molecular Diagnostic Approaches to Bacterial Infections and Antibacterial Resistance</RefTitle>
<RefYear>2018</RefYear>
<RefJournal>Annu Rev Med</RefJournal>
<RefPage>379-94</RefPage>
<RefTotal>Tsalik EL, Bonomo RA, Fowler VG Jr. New Molecular Diagnostic Approaches to Bacterial Infections and Antibacterial Resistance. Annu Rev Med. 2018 Jan 29;69:379-94. DOI: 10.1146/annurev-med-052716-030320</RefTotal>
<RefLink>https://doi.org/10.1146/annurev-med-052716-030320</RefLink>
</Reference>
<Reference refNo="13">
<RefAuthor>Korber F</RefAuthor>
<RefAuthor>Zeller I</RefAuthor>
<RefAuthor>Grünstäudl M</RefAuthor>
<RefAuthor>Willinger B</RefAuthor>
<RefAuthor>Apfalter P</RefAuthor>
<RefAuthor>Hirschl AM</RefAuthor>
<RefAuthor>Makristathis A</RefAuthor>
<RefTitle>SeptiFast versus blood culture in clinical routine - A report on 3 years experience</RefTitle>
<RefYear>2017</RefYear>
<RefJournal>Wien Klin Wochenschr</RefJournal>
<RefPage>427-34</RefPage>
<RefTotal>Korber F, Zeller I, Grünstäudl M, Willinger B, Apfalter P, Hirschl AM, Makristathis A. SeptiFast versus blood culture in clinical routine - A report on 3 years experience. Wien Klin Wochenschr. 2017 Jun;129(11-12):427-34. DOI: 10.1007/s00508-017-1181-3</RefTotal>
<RefLink>https://doi.org/10.1007/s00508-017-1181-3</RefLink>
</Reference>
<Reference refNo="14">
<RefAuthor>Quiles MG</RefAuthor>
<RefAuthor>Menezes LC</RefAuthor>
<RefAuthor>Bauab Kde C</RefAuthor>
<RefAuthor>Gumpl EK</RefAuthor>
<RefAuthor>Rocchetti TT</RefAuthor>
<RefAuthor>Palomo FS</RefAuthor>
<RefAuthor>Carlesse F</RefAuthor>
<RefAuthor>Pignatari AC</RefAuthor>
<RefTitle>Diagnosis of bacteremia in pediatric oncologic patients by in-house real-time PCR</RefTitle>
<RefYear>2015</RefYear>
<RefJournal>BMC Infect Dis</RefJournal>
<RefPage>283</RefPage>
<RefTotal>Quiles MG, Menezes LC, Bauab Kde C, Gumpl EK, Rocchetti TT, Palomo FS, Carlesse F, Pignatari AC. Diagnosis of bacteremia in pediatric oncologic patients by in-house real-time PCR. BMC Infect Dis. 2015 Jul 23;15:283. DOI: 10.1186/s12879-015-1033-6</RefTotal>
<RefLink>https://doi.org/10.1186/s12879-015-1033-6</RefLink>
</Reference>
<Reference refNo="15">
<RefAuthor>Slabisz N</RefAuthor>
<RefAuthor>Lesnik P</RefAuthor>
<RefAuthor>Zybura-Wszola K</RefAuthor>
<RefAuthor>Dudek-Wicher R</RefAuthor>
<RefAuthor>Nawrot U</RefAuthor>
<RefAuthor>Majda J</RefAuthor>
<RefTitle>Assessing the Interpretation of Molecular Test Results in the Diagnosis of Bloodstream Infections</RefTitle>
<RefYear>2024</RefYear>
<RefJournal>Diagnostics (Basel)</RefJournal>
<RefPage>915</RefPage>
<RefTotal>Slabisz N, Lesnik P, Zybura-Wszola K, Dudek-Wicher R, Nawrot U, Majda J. Assessing the Interpretation of Molecular Test Results in the Diagnosis of Bloodstream Infections. Diagnostics (Basel). 2024 Apr 27;14(9):915. DOI: 10.3390/diagnostics14090915</RefTotal>
<RefLink>https://doi.org/10.3390/diagnostics14090915</RefLink>
</Reference>
<Reference refNo="16">
<RefAuthor>Banerjee R</RefAuthor>
<RefAuthor>Teng CB</RefAuthor>
<RefAuthor>Cunningham SA</RefAuthor>
<RefAuthor>Ihde SM</RefAuthor>
<RefAuthor>Steckelberg JM</RefAuthor>
<RefAuthor>Moriarty JP</RefAuthor>
<RefAuthor>Shah ND</RefAuthor>
<RefAuthor>Mandrekar JN</RefAuthor>
<RefAuthor>Patel R</RefAuthor>
<RefTitle>Randomized Trial of Rapid Multiplex Polymerase Chain Reaction-Based Blood Culture Identification and Susceptibility Testing</RefTitle>
<RefYear>2015</RefYear>
<RefJournal>Clin Infect Dis</RefJournal>
<RefPage>1071-80</RefPage>
<RefTotal>Banerjee R, Teng CB, Cunningham SA, Ihde SM, Steckelberg JM, Moriarty JP, Shah ND, Mandrekar JN, Patel R. Randomized Trial of Rapid Multiplex Polymerase Chain Reaction-Based Blood Culture Identification and Susceptibility Testing. Clin Infect Dis. 2015 Oct 1;61(7):1071-80. DOI: 10.1093/cid/civ447</RefTotal>
<RefLink>https://doi.org/10.1093/cid/civ447</RefLink>
</Reference>
</References>
<Media>
<Tables>
<Table format="png">
<MediaNo>1</MediaNo>
<MediaID>1</MediaID>
<Caption><Pgraph><Mark1>Table 1: Primers used in this study</Mark1></Pgraph></Caption>
</Table>
<Table format="png">
<MediaNo>2</MediaNo>
<MediaID>2</MediaID>
<Caption><Pgraph><Mark1>Table 2: Identified bacteria after positive BACTEC and PCR</Mark1></Pgraph></Caption>
</Table>
<NoOfTables>2</NoOfTables>
</Tables>
<Figures>
<NoOfPictures>0</NoOfPictures>
</Figures>
<InlineFigures>
<NoOfPictures>0</NoOfPictures>
</InlineFigures>
<Attachments>
<NoOfAttachments>0</NoOfAttachments>
</Attachments>
</Media>
</OrigData>
</GmsArticle>]]></plaintext></textgroup></pgraph></abstract></origdata></gmsarticle>