Method for quickly identifying bloodstream infection pathogenic bacteria
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An identification method, a technology of pathogenic bacteria, applied in the field of microbial inspection, can solve the problems of unfavorable popularization and high price
Inactive Publication Date: 2012-05-16
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV +1
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However, sequencing and chip technology require expensive special instruments, which are expensive and are not conducive to popularization
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Embodiment 1
[0026] The identification of embodiment one positive cocci:
[0027] Gram-positive cocci on the smear;
[0028] Amplify the 16S gene fragment with 16SBF and 16SBR primers;
[0030] 20 cases of blood culture positive specimens were tested, among them, 7, 11, 16, and 18 were positive cocci. High-resolution melting curve analysis identified 2 cases of Staphylococcus aureus, 1 case of Staphylococcus epidermidis, and 1 case of Enterococcus faecalis infection.
[0031]
Embodiment 2
[0032] The identification of embodiment two negative bacilli:
[0033] smear Gram-negative bacilli;
[0034] The 16S gene fragment was amplified with 16SAF and 16SBF primers.
[0035] The common negative bacilli of 12 kinds of bacteria can be divided into 6 groups, the first group is Escherichia coli, the second group is Pseudomonas aeruginosa, Serratia marcescens, Stenotrophomonas maltophilia, and the third group is producing Enterobacter aerogenes, Klebsiella pneumoniae, Enterobacter cloacae, Klebsiella oxytoca, Burkholderia cepacia in group 4, Acinetobacter lophi in group 5, and Baumannii in group 6 Acinetobacter.
[0036] The determination of 30 cases of positive blood culture samples was completely consistent with the results obtained by subsequent traditional methods. It is confirmed that the rapid identification of pathogenic bacteria of bloodstream infection in the present invention requires only 3 hours from blood culture reporting positive, PCR amplification, and ...
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Abstract
The invention belongs to the field of examination of microorganisms, and relates to a method for quickly identifying bloodstream infection bacteria by a high-resolution fusion curve method. The method comprises the following steps of: performing smear gram staining on a blood culture positive culture flask to distinguish negative bacteria and positive bacteria; selectively amplifying different 16S fragments for the negative bacteria and the positive bacteria; and analyzing by the high-resolution fusion curve method. Common bloodstream infection pathogenic bacteria comprise staphylococcus aureus, staphylococcus epidermidis, enterococcus faecalis, enterococcus faecium, Escherichia coli, Klebsiella pneumoniae, acinetobacter baumannii, pseudomonas aeruginosa, enterobacter cloacae, Serratia marcescens, Klebsiella oxytoca, acinetobacter lwoffii, enterobacter aerogenes, stenotrophomonas maltophilia and burkholderia cepacia. By the method, the common bloodstream infection pathogenic bacteria can be measured quickly, efficiently and accurately, so that strong evidences are provided and precious time is striven for subsequent treatment.
Description
technical field [0001] The invention belongs to the field of microbiological testing, and relates to a microbiological molecular biological detection method, in particular to rapid identification of bacteria in bloodstream infection by using a high-resolution melting curve method. Background technique [0002] Bloodstream infection is a serious infectious disease, including sepsis and bacteremia. The morbidity rate is high and the case fatality rate is high. In the United States, 750,000 people develop the disease every year, of which about 215,000 people die. It is one of the top ten causes of death, accounting for 6% of all causes of death. Due to the increase in diabetes and cancer patients, the widespread use of invasive procedures, the use of hormones, chemotherapy, and drugs, and the prevalence of multidrug-resistant bacteria, the incidence of bloodstream infections has increased year by year, which has become a global problem. [0003] The traditional diagnosis ...
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