Real-time PCR strategy and detection of bacterial agents of lymphadenitis.
The aim of this study was to compare 16 S rRNA gene amplification and sequencing with a systematic real-time PCR assay screening strategy that includes all common known pathogens recovered from lymph node biopsy specimens. Lymph node biopsy samples sent to our laboratory from January 2007 to December 2008 were tested in the study. Lymph nodes were screened for the presence of any bacteria by PCR amplification and sequencing targeting the 16 S rRNA gene and also by a specific real-time PCR strategy that includes Bartonella henselae, mycobacteria, Francisella tularensis, and Tropheryma whipplei. By testing 491 lymph nodes, we found that the sensitivity of our specific real-time PCR assay strategy was significantly higher than 16 S rRNA PCR amplification and sequencing for the detection of Bartonella henselae (142 vs 98; p < 10(-4)), Francisella tularensis (16 vs 10, p < 10(-4)), and mycobacteria (8 versus 3, p < 10(-4)). None of the samples was positive for Tropheryma whipplei. Our study demonstrates the usefulness and specificity of a systematic real-time PCR strategy for molecular analysis of lymph node biopsy specimens and the higher sensitivity compared with standard 16 S rRNA gene amplification and sequencing.
Angelakis
E
E
URMITE CNRS-IRD UMR 6236, Faculté de Médecine et de Pharmacie, Université de la Méditerranée, 27 Bd Jean Moulin, 13385, Marseille Cedex 05, France.
Roux
V
V
Raoult
D
D
Rolain
J-M
JM
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001424:Bacterial Infections / Q000175:diagnosis* / Q000382:microbiology; D001431:Bacteriological Techniques / Q000379:methods*; D006801:Humans; D008199:Lymphadenitis / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S
J Clin Microbiol. 1998 Sep;36(9):2782-3
9705439
Eur Heart J. 2006 Aug;27(16):1942-6
16717077
J Clin Microbiol. 2005 Oct;43(10):5238-42
16207989
Br J Cancer. 2003 Feb 10;88(3):354-61
12569376
Clin Infect Dis. 2002 Sep 15;35(6):643-9
12203159
J Clin Microbiol. 2006 Jan;44(1):278-9
16390990
J Clin Microbiol. 1995 Jun;33(6):1573-9
7650189
Br J Surg. 1983 Jul;70(7):412-4
6871622
J Clin Microbiol. 2000 Oct;38(10):3623-30
11015374
J Clin Microbiol. 2004 Jun;42(6):2644-50
15184446
J Bacteriol. 1991 Jan;173(2):697-703
1987160
Emerg Infect Dis. 2006 Sep;12(9):1338-44
17073081
Emerg Infect Dis. 2008 Dec;14(12):1951-3
19046532
J Clin Microbiol. 2006 Mar;44(3):1018-28
16517890
Infect Dis Clin North Am. 2008 Sep;22(3):489-504, ix
18755386
Lancet Infect Dis. 2008 Mar;8(3):179-90
18291339
Medicine (Baltimore). 2008 May;87(3):167-176
18520326
Am J Med. 2002 Sep;113(4):334-6
12361821
BMC Microbiol. 2007 May 29;7:48
17535423