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Int J Infect Dis. 2018 Feb;67:122-128. doi: 10.1016/j.ijid.2017.12.015. Epub 2017 Dec 15.

Clinical utility of an optimised multiplex real-time PCR assay for the identification of pathogens causing sepsis in Vietnamese patients.

Author information

1
Department of Molecular Biology, 108 Military Central Hospital, No 1, Tran Hung Dao Street, Hai Ba Trung Dist, Hanoi, Vietnam; Vietnamese - German Centre for Medical Research (VG-CARE), Hanoi, Vietnam. Electronic address: Trungnt@benhvien108.vn.
2
Vietnamese - German Centre for Medical Research (VG-CARE), Hanoi, Vietnam; Biomedical and Pharmaceutical Applied Research Center, Vietnam Military Medical University, Hanoi, Vietnam. Electronic address: tong.van-hoang@uni-tuebingen.de.
3
Faculty of Infectious diseases, Hai Phong Medical University, 72A Nguyen Binh Khiem, Ngo Quyen District, Hai Phong, Vietnam. Electronic address: ttlien@hpmu.edu.vn.
4
Vietnamese - German Centre for Medical Research (VG-CARE), Hanoi, Vietnam; Institute of Clinical Infectious Diseases, 108 Military Central Hospital, No 1, Tran Hung Dao Street, Hai Ba Trung Dist, Hanoi, Vietnam. Electronic address: tsonntn31@gmail.com.
5
Department of Molecular Biology, 108 Military Central Hospital, No 1, Tran Hung Dao Street, Hai Ba Trung Dist, Hanoi, Vietnam. Electronic address: huyenttt@benhvien108.vn.
6
Department of Molecular Biology, 108 Military Central Hospital, No 1, Tran Hung Dao Street, Hai Ba Trung Dist, Hanoi, Vietnam. Electronic address: quyendt@benhvien108.vn.
7
Department of Molecular Biology, 108 Military Central Hospital, No 1, Tran Hung Dao Street, Hai Ba Trung Dist, Hanoi, Vietnam. Electronic address: hoanpq@benhvien108.vn.
8
Vietnamese - German Centre for Medical Research (VG-CARE), Hanoi, Vietnam; Institute of Tropical Medicine, Eberhard Karls University Tübingen, Wilhelmstr. 27, 72074, Tübingen, Germany; Faculty of Medicine, Duy Tan University, Da Nang, Vietnam. Electronic address: christian.g.meyer@gmail.com.
9
Vietnamese - German Centre for Medical Research (VG-CARE), Hanoi, Vietnam; Institute of Clinical Infectious Diseases, 108 Military Central Hospital, No 1, Tran Hung Dao Street, Hai Ba Trung Dist, Hanoi, Vietnam. Electronic address: lehuusong@108-icid.com.

Abstract

INTRODUCTION:

For the identification of bacterial pathogens, blood culture is still the gold standard diagnostic method. However, several disadvantages apply to blood cultures, such as time and rather large volumes of blood sample required. We have previously established an optimised multiplex real-time PCR method in order to diagnose bloodstream infections.

MATERIAL AND METHODS:

In the present study, we evaluated the diagnostic performance of this optimised multiplex RT-PCR in blood samples collected from 110 septicaemia patients enrolled at the 108 Military Central Hospital, Hanoi, Vietnam.

RESULTS:

Positive results were obtained by blood culture, the Light Cylcler-based SeptiFast® assay and our multiplex RT-PCR in 35 (32%), 31 (28%), and 31 (28%) samples, respectively. Combined use of the three methods confirmed 50 (45.5%) positive cases of bloodstream infection, a rate significantly higher compared to the exclusive use of one of the three methods (P=0.052, 0.012 and 0.012, respectively). The sensitivity, specificity and area under the curve (AUC) of our assay were higher compared to that of the SeptiFast® assay (77.4%, 86.1% and 0.8 vs. 67.7%, 82.3% and 0.73, respectively). Combined use of blood culture and multiplex RT-PCR assay showed a superior diagnostic performance, as the sensitivity, specificity, and AUC reached 83.3%, 100%, and 0.95, respectively. The concordance between blood culture and the multiplex RT-PCR assay was highest for Klebsiella pneumonia (100%), followed by Streptococcus spp. (77.8%), Escherichia coli (66.7%), Staphylococcus spp. (50%) and Salmonella spp. (50%). In addition, the use of the newly established multiplex RT-PCR assay increased the spectrum of identifiable agents (Acintobacter baumannii, 1/32; Proteus mirabilis, 1/32).

CONCLUSION:

The combination of culture and the multiplex RT-PCR assay provided an excellent diagnostic accomplishment and significantly supported the identification of causative pathogens in clinical samples obtained from septic patients.

KEYWORDS:

Blood culture; Bloodstream infection; Depletion of human DNA; Molecular diagnosis; Multiplex real-time PCR; Sepsis; Septicaemia

PMID:
29253706
DOI:
10.1016/j.ijid.2017.12.015
[Indexed for MEDLINE]
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