values of serum and ascetic fluid tumor necrosis factor-Alpha [TNF-alpha], TNF-receptor 1 and C-reactive protein [CRP] in patients with cirrhotic ascites

Spontaneous bacterial peritonitis [SBP] is defined as an infection of initially sterile ascitic fluid [AF] without a detectable, surgically treatable source of infection. It is a frequent and severe complication of cirrhotic ascites. Because of the high morbidity and mortality of SBP, the rapid and accurate diagnosis is required. The present study aimed to measure the levels of tumor necrosis factor-alpha [TNF-alpha], tumor necrosis factor receptor [TNF-r p55] and C-reactive protein [CRP] in both ascetic fluid and serum of patients with sterile and infected cirrhotic ascites to show their diagnostic values as compared to ascitic fluid culture and polymorphnuclear leukocyte [PMN] count. TNF-alpha, TNF-r p55 and CRP were measured in both ascetic fluid and serum of 20 patients with spontaneous bacterial peritonitis [SBP], in addition to 22 patients with sterile cirrhotic ascites. The results of clinical examination showed a significant difference as regard abdominal pain, fever jaundice, upper GIT bleedings, encephalopathy and Sclerotherapy among both groups. The serum levels of CRP and TNF-alpha were significantly higher in patients with SBP as comparing to patients have sterile ascites, but TNF-r p55 serum level showed no significant difference. On evaluation of ascetic fluid parameters, total leucocytic count [TLC], plymorphnuclear [PMN] count, CRP, TNF-alpha, TNF-r p55 are significantly higher in SBP patient group than group of sterile ascites. Sensitivity and specificity of ascitic fluid PMN [cut-off value >250 cells/ mm[3]] in discriminating infected ascites from sterile ascites were 70% and 86.4%, respectively. Sensitivity and specificity of ascitic fluid CRP [cut-off value >1.0 mg/dL] in discriminating infected ascites from sterile ascites were 85% and 72.7%, respectively. Sensitivity and specificity of ascitic fluid TNF-alpha [cut off value >12 pg/ml] in discriminating infected ascites from sterile ascites were 80% and 63.6%, respectively. Sensitivity and specificity of TNF-r p55 [cut-off value >6.2 pg/ml] in discriminating infected ascites from sterile ascites were 75% and 68.2%, respectively. We concluded that, the elevated serum and ascetic fluid levels of CRP, TNF-alpha and TNF-r may suggest their role in the pathogenesis of ascetic fluid infection and their higher sensitivity and specificity make them to be good discriminators in ascetic fluid infection [especially a cheap and easy ascitic fluid CRP levels]. Thus may help in rapid diagnosis and early start empirical antibiotic therapy without waiting the culture results

clinical significance of real-time PCR for rapid diagnosis of mycoplasma pneumoniae in respiratory tract infection

Mycoplasma pneumoniae [M. pneumoniae] is a common cause of respiratory tract infections [RTIs]. Diagnosis of M. pneumoniae infection relies mainly on laboratory tests, as the clinical presentation is not significantly different from that seen with other pathogens causing RTI. Diagnosis has traditionally been obtained by serological diagnosis, but increasingly, molecular techniques have been applied. However, the number of studies actually comparing these assays is limited. The Real Time PCR [RT-PCR] assay for detection of M. pneumoniae was used and comparing with a conventional PCR assay, and with serology using IgM Immunofluorescence assay [IFA]. The study included: 70 adult patients with manifestations of RTIs and 20 age matched healthy controls. All patients and controls subjected to the following: thorough history and clinical examination, routine investigations as complete blood count [CBC], erythrocyte sedimentation rate [ESR], C-reactive protein [CRP], and chest-X-ray, blood samples were collected for serological testing [IgM-IFA]. nasopharyngeal swabs were done for PCR assays and for culture techniques. Diagnosis of M. pneumoniae was made on the basis of tbe conventional PCR results. The conventional PCR was positive for M. pneumoniae in 18 patients [25.7%] out of overall 70 patients and all the 18 patients positive by conventional PCR were also positive by RT-PCR. While, all controls were PCR negative by both techniques. The nasopharyngeal swab culture for M. pneumoniae was positive in 10[14.3%] out of 70 patients and negative in all 20 healthy controls. However, the M. pneumoniae IgM-IFA was positive for 21 [30%] out of 70 patients. So IgM IFA was positive in 3 patients with negative PCR, However, one case was positive for IgM- IFA among healthy controls. Comparing to conventional PCR, the RT-PCR gives 100% sensitivity, 100% specificity and 100% accuracy for diagnosis of M. pneumoniae, followed by IgM- IFA that give a sensitivity of 100%, specificity of 94.2% and accuracy of 95.7%, and the culture of M. pneumoniae gives 55.6% for sensitivity, 100% for specificity and 88.5% for diagnostic accuracy. The comparison of clinical data of patients diagnosed with M. pneumoniae infection [+ve PCR] and those who were M. pneumoniae negative [-ye PCR] revealed that no significant difference was reported as regard age or disease duration between the two groups of patients, while rhinitis was significantly more prevalent in the Mycoplasma-negative patients [p<0.01]. No significant difference was seen between the two groups regarding headache, malaise, sore throat, vomiting, fever, wheeze, or chest radiographic infiltrate [p>0.05]. Also, WBC counts, CRP levels and ESR values were significantly increased in PCR positive group as comparing to PCR negative group [p<0.05]. The molecular methods are superior for diagnosis of M. pneumoniae regarding accuracy, and providing more rapid diagnosis. In addition, using Real-Time PCR assay involves less hands, short time for diagnosis and meanwhile, rapid treatment and monitoring therapy