Expression profiles for predicting septic conditions
A rapid, safe method for predicting sepsis, or a condition similar to sepsis, in a mammal is disclosed, which comprises the following steps: I) isolating RNA from a biological sample of a mammal; ii) labeling the isolated RNA of step I) with a detectable marker; iii) hybridizing the labeled isolated RNA of step ii) with at least one DNA of genes Seq.-ID 1 to Seq.-ID 6705 (Table 1), which is spotted onto a microarray and which is a sepsis-specific gene or gene fragment, under the reaction conditions for hybridizations; iv) quantitatively recording labeling signals of the hybridized RNA of step iii) in an expression profile; v) comparing the expression profile of step iv) with a control sample with respect to a stronger or weaker expression of genes or gene fragments which are specific for sepsis; and vi) combining the expression profile of step v) with protein- and metabolite patterns of the biological sample.
1. A method for in-vitro detection of sepsis in a blood sample of a human inpatient selected from the group consisting of a polytraumatized adult patient, an adult patient with pneumonia, and a prematurely born infant, the method comprising the following steps:
i) extracting RNA from a blood sample of the human inpatient;
ii) quantifying the RNA extracted during step (i), and transcribing and labeling the RNA extracted during step (i) to obtain a quantity of labeled cRNA, wherein for labeling a detectable marker is used, wherein said detectable marker is a fluorescence marker, dye marker or a radioactive marker;
iii) fragmenting the labeled cRNA obtained according to step (ii) , and specifically hybridizing the labeled, fragmented cRNA, if present with polynucleotides on a microarray comprising DNA which corresponds to polynucleotides SEQ ID NO: 2 and SEQ ID NO: 6704 (Table 1) to obtain labeled, fragmented, hybridized cRNA;
iv) obtaining control labeled and fragmented cRNA from a human with no sepsis wherein said control labeled fragmented cRNA is obtained in accordance with steps (i), (ii) and (iii), and hybridizing a known quantity of the control labeled, fragmented cRNA to said microarray in accordance with step (iii);
v) determining a quantitative expression profile for both the human inpatient and the control labeled cRNA; and
vi) comparing the expression profiles of step (v) of the human inpatient and of the control wherein statistically significant overexpression of the gene corresponding to SEQ ID NO: 2 and statistically significant underexpression of the gene corresponding to SEQ ID NO: 6704 for the human inpatient relative to the control is indicative of the patient's having sepsis.
2. A DNA-microarray for diagnosis of sepsis in a blood sample of a human, said microarray consisting of at least one of the following groups:
SEQ ID NO: 6706 to SEQ ID NO: 7562 (Table 2),
SEQ ID NO: 7563 to SEQ ID NO: 8947 (Table 3)
SEQ ID NO: 11379 to SEQ ID NO: 13905 (Table 7), and
SEQ ID NO: 13906 to SEQ ID NO: 14075 (Table 8),
wherein said oligonucleotides are the only oligonucleotides forming the microarray.