Early detection of malignant showers in blood of colorectal cancer patients using carcinoembryonic antigen mRNA Reverse Transcriptase-polymerase Chain Reaction

Serum CEA is among the most widely accepted marker for diagnosing and monitoring colorectal cancer [CRC] yet it has many limitations. The objective of the present study was to evaluate the utility of plasma CEA-mRNA as a marker for early detection of micrometastasis and assess its usefulness versus that of serum CEA in CRC patients. This study included 36 patients with CRC who were staged according to Dukes' staging system into stage A [n = 4], B [n = 8] C [n = 14] and stage D [n = 10]. Patients included in Dukes' stages A and B [n = 12] had non-metastatic lesion and were considered as one group [group I] while those having stages C and D [n = 24] had metastatic lesions. Patients with metastatic lesions who weren't receiving chemotherapy were included in one group [group II, n = 20] while those on chemotherapy [5 fluorouracil and leukovorin] were considered as group III [n = 4]. Six patients suffering from benign colorectal disease [ulcerative colitis n = 2, diverticulitis n = 1 and polyps n=3] and six healthy age and gender matched and with normal serum CEA concentration were included in the study and were considered as the control group [group IVa, group IVb respectively]. All CRC patients were subjected to clinical, radiological, endoscopic, histopathological and laboratory assessment. Control subjects were assessed both clinically and laboratory wise. Serum CEA was assayed by chemiluminescent sequential immunometric assay while plasma CEA-mRNA was determined by semi nested reverse transcription RT-PCR. The median and inter-quartile values for serum CEA in groups I, II and III were 2.5 [1.5-3.4], 4.6 [2.8-9.9] and 4.2 [2.9-5.3] ng/mL respectively. As for the control groups, it was 2 [0.9-2.2] ng/mL in group IVa and 3.3 [2.1-4.0] ng/mL in group IVb. When compared to controls, serum CEA was significantly higher only in metastatic patients [group II and III], however, when compared to non-metastatic CRC, serum CEA was significantly higher only in patients not on chemotherapy [group II]. The degree of tumor differentiation had no significant impact on serum CEA concentration. Our results also demonstrated that the percent positivity for CEA-mRNA increase with advancing CRC stage where a highly significant increase was observed in metastatic patients [65%] when compared to non metastatic ones [8.3%]. Our results also showed that CEA- mRNA may be positive in subjects with benign colorectal diseases [8.3%] and that chemotherapy may result in negative results for m-RNA. Again, the degree of differentiation had no impact on percent positivity of CEA- mRNA. Finally, our results showed that serum CEA and percent positivity of CEA- mRNA do not correlate and that the recurrence rate in patients with positive CEA-mRNA is significantly high [70%]. Reverse-trancriptase polymerase chain reaction for CEA-mRNA is a sensitive method for detection of circulating cancer cells in CRC patients. Colorectal cancer patients with postoperative CEA-mRNA positive cells in peripheral blood have less disease free survival than patients who demonstrate absence of these cells. Studies involving a larger group of patients with a longer follow-up period should be done to implement the clinical relevance of this phenomenon. Patients receiving chemotherapy should not be tested for CEA-mRNA during the treatment course. They should be tested repeatedly thereafter at longer time intervals following the last dose of chemotherapy

Plasma matrix metalloproteinase 1 [MMP1] and tissue inhibitor of metalloproteinase 1 [TIMP1]: potential diagnostic markers of prostate cancer progression

The purpose of this work was to evaluate the role of plasma matrix metalloproteinase 1 [MMP1] and tissue inhibitor of metalloproteinase 1 [TIMP1] in diagnosis and staging of prostate cancer patients. This study was conducted on 60 male patients [aged 48-80 years] and 20 age matched healthy individuals. Patients were classified according to clinical examination, prostate specific antigen [PSA] findings, pathological and radiological findings into : 25 patients with benign prostatic hyperplasia [BPH] and 35 patients with different stages of prostate cancer [PC] divided into : [13 PC with no metastasis, 13 PC with local metastasis and 9 PC with distant metastasis]. Plasma MMP1 and TIMP1 were assayed using sandwich immunoassay technique for all subjects. Serum total prostate specific antigen [TPSA] and serum alkaline phosphatase [ALP] were measured to all patients and control group. Free PSA and PSA index were performed for patients with TPSA between 4 and 10 ng/mL. PSA index was found to be superior to TPSA in discriminating patients with prostate cancer [p < 0.001]. There was no significant difference in plasma MMPI levels between different studied groups [p > 0.05]. Serum ALP was significantly increased in PC patients with distant metastasis than those with local metastasis; ALP was additionally found to be the best predictive marker for the presence of distant bone metastasis. Using ROC, Plasma ALP showed a diagnostic sensitivity for bone metastasis in prostate cancer patients of 100% at a cutoff of 90IU/L Median levels of plasma TIMP1 were found to be increased in PC patients [n = 35] over BPH [n = 25] [1600, 180 ug/L respectively]. Additionally, plasma TIMP1 median levels showed a progressive increase with malignant progression of PC being 400 ug/L in PC with no metastasis [n = 13], 2200 ug/L in PC with local metastasis [n = 13] and 4000 ug/L in PC with distant bone metastasis [n = 9], with a statistically significant difference between groups [p < 0.001]. There was a significant negative correlation between TIMP1 and PSA index in PC patients. TIMP1 also correlated positively with ALP in the same group. Mean-while, TIMP1 proved to best discriminate PC patients with metastasis [local and distant] from those with no metastasis. Accordingly, plasma TIMP1 is accurately correlated with staging of prostate cancer and is a promising marker of malignant progression of prostate cancer. Moreover, addition of ALP can effectively detect distant bone metastasis in PC patients

clinical utility of ELISA method for diagnosis in children

This study was conducted on 42 patients [22 males and 20 females] suffering from symptoms suggestive of giardiasis and 12 age and sex matched control subjects aiming to assess clinical utility of enzyme- linked immunosorbent assay [ELISA] for the detection of Giardia antigen in stool specimens of patients. The results revealed a much superior diagnostic performance of ELISA method compared with routine microscopic examination of stool specimens for the detection of the parasite. ELISA method showed 97.6% sensitivity compared with 83.3% in case of microscopic examination of stool specimen. In addition, ELISA method proved to be 100% specific yielding a diagnostic accuracy of 96.4%. The predictive value of a positive test was 100%, whereas that of a negative test was 92.3%. Unlike microscopic examination of duodenal aspirate samples, ELISA method for the detection of Giardia antigen in stool specimens proved to be a simple test which can be easily performed on a single stool specimen yielding reliable results that ensure prompt initiation of a proper line treatment and avoidance of blind treatment regimens