Use of a DNA probe to detect Salmonella typhi in the blood of patients with typhoid fever

Rubin, Fran A.; McWhirter, Paul D.; Punjabi, N H; Lane, Edward M.; Sudarmono, Pratiwi; Pulungsih, Sri Pandam; Lesmana, Murad; Kumala, Swiandy; Kopecko, Dennis J.; Hoffman, Stephen L.

doi:10.1128/jcm.27.5.1112-1114.1989

Cited by 29 publications

(4 citation statements)

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“…In the present study severely ill patients, who comprise less than 5% of all cases in this area, were not included (although 14 of 374 patients did have evidence of upper gastrointestinal bleeding). The range of bacterial counts in blood in this large study of typhoid fever is similar to those in the much-smaller studies reported previously by Rubin et al, using first a semiquantitative DNA hybridization method (19) and, second, direct cultures (18). In terms of intrinsic toxicity there is little difference between enterobacteriaceae.…”

Section: Discussionsupporting

confidence: 87%

Quantitation of Bacteria in Blood of Typhoid Fever Patients and Relationship between Counts and Clinical Features, Transmissibility, and Antibiotic Resistance

et al. 1998

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Salmonella typhi was isolated from 369 andSalmonella paratyphi A was isolated from 6 of 515 Vietnamese patients with suspected enteric fever. Compared with conventional broth culture of blood, direct plating of the buffy coat had a diagnostic sensitivity of 99.5% (95% confidence interval [CI], 97.1 to 100%). Blood bacterial counts were estimated by the pour plate method. The median S. typhi count in blood was 1 CFU/ml (range, <0.3 to 387 CFU/ml), of which a mean of 63% (95% CI, 58 to 67%) were intracellular. The mean number of bacteria per infected leukocyte was 1.3 (interquartile range [IQR], 0.7 to 2.4) CFU/cell (n = 81). Children (<15 years old;n = 115) had higher median blood bacterial counts than adults (n = 262): 1.5 (range, <0.3 to 387) versus 0.6 (range, <0.3 to 17.7) CFU/ml (P = 0.008), and patients who excreted S. typhi in feces had higher bacteremias than those who did not: a median of 3 (range, <0.3 to 32) versus 1 (range, <0.3 to 68) CFU/ml (P = 0.02). Blood bacterial counts declined with increasing duration of illness (P = 0.002) and were higher in infections caused by multidrug-resistant S. typhi (1.3 [range, <0.3 to 387] CFU/ml; n = 313) than in infections caused by antibiotic-sensitive S. typhi (0.5 [range, <0.3 to 32] CFU/ml; n = 62) (P = 0.006). In a multivariate analysis this proved to be an independent association, suggesting a relationship between antibiotic resistance and virulence in S. typhi.

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Section: Discussionsupporting

confidence: 87%

Quantitation of Bacteria in Blood of Typhoid Fever Patients and Relationship between Counts and Clinical Features, Transmissibility, and Antibiotic Resistance

et al. 1998

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“…The low concentration of S. typhi cells in the blood of patients with typhoid fever, <15 bacteria per ml (9,11), undoubtedly contributes to the moderate sensitivity of blood culture and antigen detection tests. We speculated that if all bacteria in a blood sample could be concentrated, this would facilitate culture of the organism and detection of bacterial antigens and DNA.…”

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Rapid diagnosis of typhoid fever through identification of Salmonella typhi within 18 hours of specimen acquisition by culture of the mononuclear cell-platelet fraction of blood

et al. 1990

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Detection of Salmonella typhi in blood by culture of the mononuclear cell-platelet layer was compared with other methods currently used for the diagnosis of typhoid fever. Colonies of S. typhi were present in all mononuclear cell-platelet layer-positive cultures within 18 h of plating and were identified within an additional 10 min by a coagglutination technique. In contrast, identification of all positive cultures by conventional blood culture required 3 days.

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“…Rapid diagnosis can be achieved by the direct detection of characteristic bacterial genes in clinical specimens, and many primer sets have been developed to detect species-specific genes in simple PCRs (10,18,31). These systems are usually designed to confirm the diagnosis of specific clinical syndromes and include the identification of Burkholderia pseudomallei in melioidosis (9,34), S. pneumoniae in pneumococcal pneumonia and meningitis (7,22), Coxiella burnettii in Q fever (46), Listeria monocytogenes in listeriosis (5), Rhodococcus equi in rhodococcosis in horses (42), Mycobacterium tuberculosis in tuberculosis (12,36), Salmonella enterica serovar Typhi in typhoid fever (38,44), Mycoplasma pneumoniae in mycoplasmal pneumonia (30), Neisseria meningitidis in meningococcal meningitis (31), and Borrelia burgdorferi in Lyme disease (15). Similar test systems have been designed for yeasts (29).…”

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Rapid Diagnosis of Bacteremia by Universal Amplification of 23S Ribosomal DNA Followed by Hybridization to an Oligonucleotide Array

2000

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The rapid identification of bacteria in blood cultures and other clinical specimens is important for patient management and antimicrobial therapy. We describe a rapid (<4 h) detection and identification system that uses universal PCR primers to amplify a variable region of bacterial 23S ribosomal DNA, followed by reverse hybridization of the products to a panel of oligonucleotides. This procedure was successful in discriminating a range of bacteria in pure cultures. When this procedure was applied directly to 158 unselected positive blood culture broths on the day when growth was detected, 125 (79.7%) were correctly identified, including 4 with mixed cultures. Nine (7.2%) yielded bacteria for which no oligonucleotide targets were present in the oligonucleotide panel, and 16 culture-positive broths (10.3%) produced no PCR product. In seven of the remaining eight broths, streptococci were identified but not subsequently grown, and one isolate of Staphylococcus aureus was misidentified as a coagulase-negative staphylococcus. The accuracy, range, and discriminatory power of the assay can be continually extended by adding further oligonucleotides to the panel without significantly increasing complexity or cost.

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Use of a DNA probe to detect Salmonella typhi in the blood of patients with typhoid fever

Cited by 29 publications

References 10 publications

Quantitation of Bacteria in Blood of Typhoid Fever Patients and Relationship between Counts and Clinical Features, Transmissibility, and Antibiotic Resistance

Quantitation of Bacteria in Blood of Typhoid Fever Patients and Relationship between Counts and Clinical Features, Transmissibility, and Antibiotic Resistance

Rapid diagnosis of typhoid fever through identification of Salmonella typhi within 18 hours of specimen acquisition by culture of the mononuclear cell-platelet fraction of blood

Rapid Diagnosis of Bacteremia by Universal Amplification of 23S Ribosomal DNA Followed by Hybridization to an Oligonucleotide Array

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