Diagnostic utility of in-house loop-mediated isothermal amplification and real-time PCR targeting virB gene for direct detection of Brucella melitensis from clinical specimens

S. Patra, C. Tellapragada, K. E. Vandana, C. Mukhopadhyay

Research output: Contribution to journalArticle

Abstract

Aims: In this present study, the utility of a newly developed loop-mediated isothermal amplification (LAMP) and real-time PCR assays designed to amplify the virB gene region of Brucella melitensis was evaluated from human clinical specimens. Methods and Results: Fifty-four culture-confirmed cases of brucellosis and 54 culture negative but clinically suspected cases of brucellosis were included in the study. Whole blood, serum and other nonblood specimens were collected and subjected to blood culture using automatic blood culture system, serological tests, LAMP assay and real-time PCR. Overall sensitivities of LAMP and real-time PCR assays were 67·5 and 68·3% respectively. For nonblood clinical specimens, we noticed a marked increase in the sensitivities of LAMP (88·9%) and real-time PCR (100%) assays. Conclusions: Performance of LAMP and real-time PCR was not satisfactory for whole-blood specimens because of the low abundance of bacteria or DNA. On the other hand, using nonblood specimens, both the assays showed higher sensitivity and specificity which makes them a good alternative for the rapid diagnosis of human brucellosis. Significance and Impact of the Study: The developed LAMP and real-time PCR assays are a specific and rapid diagnostic tool for direct and early detection of Brucella in clinical specimens.

Original languageEnglish
Pages (from-to)230-236
Number of pages7
JournalJournal of Applied Microbiology
Volume127
Issue number1
DOIs
Publication statusPublished - 01-07-2019

Fingerprint

Brucella melitensis
Gene Targeting
Real-Time Polymerase Chain Reaction
Brucellosis
Brucella
Serologic Tests
Bacteria
Sensitivity and Specificity
DNA
Serum
Genes

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

@article{3991e01e603645aa84a6c0ff169cf88f,
title = "Diagnostic utility of in-house loop-mediated isothermal amplification and real-time PCR targeting virB gene for direct detection of Brucella melitensis from clinical specimens",
abstract = "Aims: In this present study, the utility of a newly developed loop-mediated isothermal amplification (LAMP) and real-time PCR assays designed to amplify the virB gene region of Brucella melitensis was evaluated from human clinical specimens. Methods and Results: Fifty-four culture-confirmed cases of brucellosis and 54 culture negative but clinically suspected cases of brucellosis were included in the study. Whole blood, serum and other nonblood specimens were collected and subjected to blood culture using automatic blood culture system, serological tests, LAMP assay and real-time PCR. Overall sensitivities of LAMP and real-time PCR assays were 67·5 and 68·3{\%} respectively. For nonblood clinical specimens, we noticed a marked increase in the sensitivities of LAMP (88·9{\%}) and real-time PCR (100{\%}) assays. Conclusions: Performance of LAMP and real-time PCR was not satisfactory for whole-blood specimens because of the low abundance of bacteria or DNA. On the other hand, using nonblood specimens, both the assays showed higher sensitivity and specificity which makes them a good alternative for the rapid diagnosis of human brucellosis. Significance and Impact of the Study: The developed LAMP and real-time PCR assays are a specific and rapid diagnostic tool for direct and early detection of Brucella in clinical specimens.",
author = "S. Patra and C. Tellapragada and Vandana, {K. E.} and C. Mukhopadhyay",
year = "2019",
month = "7",
day = "1",
doi = "10.1111/jam.14260",
language = "English",
volume = "127",
pages = "230--236",
journal = "Journal of Applied Microbiology",
issn = "1364-5072",
publisher = "Wiley-Blackwell",
number = "1",

}

TY - JOUR

T1 - Diagnostic utility of in-house loop-mediated isothermal amplification and real-time PCR targeting virB gene for direct detection of Brucella melitensis from clinical specimens

AU - Patra, S.

AU - Tellapragada, C.

AU - Vandana, K. E.

AU - Mukhopadhyay, C.

PY - 2019/7/1

Y1 - 2019/7/1

N2 - Aims: In this present study, the utility of a newly developed loop-mediated isothermal amplification (LAMP) and real-time PCR assays designed to amplify the virB gene region of Brucella melitensis was evaluated from human clinical specimens. Methods and Results: Fifty-four culture-confirmed cases of brucellosis and 54 culture negative but clinically suspected cases of brucellosis were included in the study. Whole blood, serum and other nonblood specimens were collected and subjected to blood culture using automatic blood culture system, serological tests, LAMP assay and real-time PCR. Overall sensitivities of LAMP and real-time PCR assays were 67·5 and 68·3% respectively. For nonblood clinical specimens, we noticed a marked increase in the sensitivities of LAMP (88·9%) and real-time PCR (100%) assays. Conclusions: Performance of LAMP and real-time PCR was not satisfactory for whole-blood specimens because of the low abundance of bacteria or DNA. On the other hand, using nonblood specimens, both the assays showed higher sensitivity and specificity which makes them a good alternative for the rapid diagnosis of human brucellosis. Significance and Impact of the Study: The developed LAMP and real-time PCR assays are a specific and rapid diagnostic tool for direct and early detection of Brucella in clinical specimens.

AB - Aims: In this present study, the utility of a newly developed loop-mediated isothermal amplification (LAMP) and real-time PCR assays designed to amplify the virB gene region of Brucella melitensis was evaluated from human clinical specimens. Methods and Results: Fifty-four culture-confirmed cases of brucellosis and 54 culture negative but clinically suspected cases of brucellosis were included in the study. Whole blood, serum and other nonblood specimens were collected and subjected to blood culture using automatic blood culture system, serological tests, LAMP assay and real-time PCR. Overall sensitivities of LAMP and real-time PCR assays were 67·5 and 68·3% respectively. For nonblood clinical specimens, we noticed a marked increase in the sensitivities of LAMP (88·9%) and real-time PCR (100%) assays. Conclusions: Performance of LAMP and real-time PCR was not satisfactory for whole-blood specimens because of the low abundance of bacteria or DNA. On the other hand, using nonblood specimens, both the assays showed higher sensitivity and specificity which makes them a good alternative for the rapid diagnosis of human brucellosis. Significance and Impact of the Study: The developed LAMP and real-time PCR assays are a specific and rapid diagnostic tool for direct and early detection of Brucella in clinical specimens.

UR - http://www.scopus.com/inward/record.url?scp=85067051007&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85067051007&partnerID=8YFLogxK

U2 - 10.1111/jam.14260

DO - 10.1111/jam.14260

M3 - Article

C2 - 30897267

AN - SCOPUS:85067051007

VL - 127

SP - 230

EP - 236

JO - Journal of Applied Microbiology

JF - Journal of Applied Microbiology

SN - 1364-5072

IS - 1

ER -