The Study Of Bacterial PCR Combined With Procalcitonin In Diagnosis Of Acute Pancreatitis Complicated With Systemic Infections

Objective:To explore the diagnostic value of 16SrRNA and 18SRrNA genes based on bacteria and fungi PCR method in diagnosing acute paneratitis complicated with systemic infections. Represented the characteristics of the pathogen infection.To explore a method for rapid detection of bacterial infection.Methods:97 patients with acute pancrcatitis(during 2009-9 to 2010-12) was randomly collected(Male 51 cases, female 25 cases, aged 14-72).106 samples came from different periods and parts(sputum 12 cases,ascites 18 cases,blood 76 cases).Patients were included on the basis of the guide of China's treatment of acute pancreatitis (2007).Firstly,a universal primer(UP)was synthesized based on the highly conserved sequences of bacterial 16SrRNA and fungi 18SrRNA genes.106 samples were DNA extracted and amplified by polymerase chain reaction(PCR),Results were observed with agarose gel electrophoresis(AGE).The specificity and sensitivity of the PCR were tested.PCR detection was carried out to compare with conventional culture results.SPSS11.5 for Windows software was used to carry out statistical treatment.Resluts:For all bacterial PCR,the positive fragment with length of 1500bp were amplified from all strains. No signal was observed when human DNA and viruses were used as templates.The primer can only amplifiy bacteria and its sensitivity could be improved to 10CFU/ml E coli. And for all fungi PCR positive fragment with length of 197bp were amplified from all strains. No signal was observed when human DNA and bacteria were used as templates. Its low limits to 10CFU/ml Candida mycoderma.Different types of 106 specimens,the total positive rate on culture is 20.75%(22/106).Among them,the positive rate of Bacteria culture is 19.81%(20/106) and fungi culture are is 0.94%(1/106);The total positive rate of PCR is 32.07%(34/106).Of the total,the positive rate of bacteria 16SrRNA gene PCR are 28.30%(30/106) and fungi 18SrRNA gene PCR is 3.77%(4/106);the consistent positive rate of culture and PCR is 18.87%(20/106).PCR was significantly higher than that culture (P<0.05),and the sensitivity is 90.9%(20/22),the specificity is 83.33%(70/84).PCR test generally takes 6-8 hours, while the traditional culturvation method will need 3 days.The most commonly infected site is abdominal cavity,secondly is respiratory tract and blood.Conclusion:16SrRNA and 18SrRNA genes based on bacteria and fungi UP-PCR method can quickly diagnosing acute pancreatitis complicated with systemic infections and have high sensitivity and specificity.The bacteria primarily are E coli and Acinetobacter baumannii.Part 2:The Value Of Procalcitonin In Diagnosis Of Acute Pancreatitis Complicated With Early Infection AbstractObjective:To evaluate the clinical diagnostic value of detect procalcitonin's expression in acute pancreatitis complicated with infections.Methods:76 patients with acute pancrcatitis was randomly selected in first part of patients.According to whether or not complicating with systemic inflammatory response syndrome(SIRS),patients were divided into SIRS groups and negative SIRS groups(15 cases).On foundation of infection SIRS groups were divided into SIRS groups(53 cases) and SIRS infection groups(8 cases).Serum from patients was measured on the first week after admission, Enzyme—Linked Immunosorbent Assay(ELISA)was used to examine the expression of PCT。SPSS 11.5 for Windows software was used to carry out statistical treatment.And according to the first part of PCR detection results,The predictive power of PCT and CRP was compared in patients of cute pancreatitis complicated with infections.Draw the receiver characteristic curve(ROC curve).Resluts:The results of blood serum PCT quantitate Enzyme—Linked immunodetection were displayed that Serum PCT level of SIRS infection groups (2.68±0.84) ng/ml are obviously higher than that negative SIRS groups(0.97±0.56) ng/ml and SIRS groups(1.39±0.58) ng/ml (P<0.05).But there are no significant differences of CRP in three groups(P>0.05).According to the first part of PCR detection results,Draw the receiver characteristic curve(ROC curve).The predicte infections power of PCT(AUC=0.737,P=0.001) is stronger than CRP(AUC=0.703, P=0.004)。For 2.0ng/ml as predict threshold,The sensitivity and specificity was 74.1% and 67.5% respectively.Conclusion:PCT is a better diagnosis and observing indexes in acute pancreatitis complicated with early infection.