Cargando…

Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for rapid diagnosis of neonatal sepsis

BACKGROUND & OBJECTIVES: The difficulties in diagnosis of neonatal sepsis are due to varied clinical presentation, low sensitivity of blood culture which is considered the gold standard and empirical antibiotic usage affecting the outcome of results. Though polymerase chain reaction (PCR) based...

Descripción completa

Detalles Bibliográficos
Autores principales: Rohit, Anusha, Maiti, Biswajit, Shenoy, Shalini, Karunasagar, Indrani
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Medknow Publications & Media Pvt Ltd 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4822372/
https://www.ncbi.nlm.nih.gov/pubmed/26997017
http://dx.doi.org/10.4103/0971-5916.178613
_version_ 1782425774708490240
author Rohit, Anusha
Maiti, Biswajit
Shenoy, Shalini
Karunasagar, Indrani
author_facet Rohit, Anusha
Maiti, Biswajit
Shenoy, Shalini
Karunasagar, Indrani
author_sort Rohit, Anusha
collection PubMed
description BACKGROUND & OBJECTIVES: The difficulties in diagnosis of neonatal sepsis are due to varied clinical presentation, low sensitivity of blood culture which is considered the gold standard and empirical antibiotic usage affecting the outcome of results. Though polymerase chain reaction (PCR) based detection of bacterial 16S rRNA gene has been reported earlier, this does not provide identification of the causative agent. In this study, we used restriction fragment length polymorphism (RFLP) of amplified 16S rRNA gene to identify the organisms involved in neonatal sepsis and compared the findings with blood culture. METHODS: Blood samples from 97 neonates were evaluated for diagnosis of neonatal sepsis using BacT/Alert (automated blood culture) and PCR-RFLP. RESULTS: Bacterial DNA was detected by 16S rRNA gene PCR in 55 cases, while BacT/Alert culture was positive in 34 cases. Staphylococcus aureus was the most common organism detected with both methods. Klebsiella spp. was isolated from four samples by culture but was detected by PCR-RFLP in five cases while Acinetobacter spp. was isolated from one case but detected in eight cases by PCR-RFLP. The sensitivity of PCR was found to be 82.3 per cent with a negative predictive value of 85.7 per cent. Eighty of the 97 neonates had prior exposure to antibiotics. INTERPRETATION & CONCLUSIONS: The results of our study demonstrate that PCR-RFLP having a rapid turnaround time may be useful for the early diagnosis of culture negative neonatal sepsis.
format Online
Article
Text
id pubmed-4822372
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Medknow Publications & Media Pvt Ltd
record_format MEDLINE/PubMed
spelling pubmed-48223722016-04-25 Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for rapid diagnosis of neonatal sepsis Rohit, Anusha Maiti, Biswajit Shenoy, Shalini Karunasagar, Indrani Indian J Med Res Original Article BACKGROUND & OBJECTIVES: The difficulties in diagnosis of neonatal sepsis are due to varied clinical presentation, low sensitivity of blood culture which is considered the gold standard and empirical antibiotic usage affecting the outcome of results. Though polymerase chain reaction (PCR) based detection of bacterial 16S rRNA gene has been reported earlier, this does not provide identification of the causative agent. In this study, we used restriction fragment length polymorphism (RFLP) of amplified 16S rRNA gene to identify the organisms involved in neonatal sepsis and compared the findings with blood culture. METHODS: Blood samples from 97 neonates were evaluated for diagnosis of neonatal sepsis using BacT/Alert (automated blood culture) and PCR-RFLP. RESULTS: Bacterial DNA was detected by 16S rRNA gene PCR in 55 cases, while BacT/Alert culture was positive in 34 cases. Staphylococcus aureus was the most common organism detected with both methods. Klebsiella spp. was isolated from four samples by culture but was detected by PCR-RFLP in five cases while Acinetobacter spp. was isolated from one case but detected in eight cases by PCR-RFLP. The sensitivity of PCR was found to be 82.3 per cent with a negative predictive value of 85.7 per cent. Eighty of the 97 neonates had prior exposure to antibiotics. INTERPRETATION & CONCLUSIONS: The results of our study demonstrate that PCR-RFLP having a rapid turnaround time may be useful for the early diagnosis of culture negative neonatal sepsis. Medknow Publications & Media Pvt Ltd 2016-01 /pmc/articles/PMC4822372/ /pubmed/26997017 http://dx.doi.org/10.4103/0971-5916.178613 Text en Copyright: © Indian Journal of Medical Research http://creativecommons.org/licenses/by-nc-sa/3.0 This is an open access article distributed under the terms of the Creative Commons Attribution NonCommercial ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non commercially, as long as the author is credited and the new creations are licensed under the identical terms.
spellingShingle Original Article
Rohit, Anusha
Maiti, Biswajit
Shenoy, Shalini
Karunasagar, Indrani
Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for rapid diagnosis of neonatal sepsis
title Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for rapid diagnosis of neonatal sepsis
title_full Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for rapid diagnosis of neonatal sepsis
title_fullStr Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for rapid diagnosis of neonatal sepsis
title_full_unstemmed Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for rapid diagnosis of neonatal sepsis
title_short Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) for rapid diagnosis of neonatal sepsis
title_sort polymerase chain reaction-restriction fragment length polymorphism (pcr-rflp) for rapid diagnosis of neonatal sepsis
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4822372/
https://www.ncbi.nlm.nih.gov/pubmed/26997017
http://dx.doi.org/10.4103/0971-5916.178613
work_keys_str_mv AT rohitanusha polymerasechainreactionrestrictionfragmentlengthpolymorphismpcrrflpforrapiddiagnosisofneonatalsepsis
AT maitibiswajit polymerasechainreactionrestrictionfragmentlengthpolymorphismpcrrflpforrapiddiagnosisofneonatalsepsis
AT shenoyshalini polymerasechainreactionrestrictionfragmentlengthpolymorphismpcrrflpforrapiddiagnosisofneonatalsepsis
AT karunasagarindrani polymerasechainreactionrestrictionfragmentlengthpolymorphismpcrrflpforrapiddiagnosisofneonatalsepsis