applicability of T-cell receptor gamma gen rearrangement as an adjuvant diagnosis tool in skin biopsies for cutaneous T-cell lymphoma

The diagnosis of cutaneous T-cell lymphoid infiltrates may be difficult based on clinical and routine immunohistologic findings. In this situation, an ancillary technique demonstrating the presence of a monoclonal cell proliferation could help to rule in or out cutaneous T-cell lymphoma [CTCL] in cases that clinically and histopathologically do not allow a definitive diagnosis. Southern blot analysis is a time-consuming method with low sensitivity that should not be considered for the routine diagnosis of cutaneous lymphoid infiltrates. Moreover, it can be used only when fresh tissue is available. New assays based on the amplification of the T-cell receptor gamma [TCR gamma] chain gene rearrangement by polymerase chain reaction [PCR] have been proposed to overcome these limitations. We retrospectively studied 124 biopsies from 104 patients [66 biopsies with the clinical and histological diagnosis or suspicious of CTCL and 58 biopsies with histological diagnosis of benign reactive dermatological conditions who presented to the Dermatology Unit at King Faisal Specialist Hospital and Research Center, Riyadh, Kingdom of Saudi Arabia between 1996 and 2004. The specimens were morphologically examined and then analyzed by PCR for the gamma chain of the TCR gamma followed by gel electrophoresis. The results showed 87.1% sensitivity and 92% specificity in detecting clonal T-cell gene rearrangements among CTCL cases with a positive predictive value of 93.1% and negative predictive value of 85.2%. Therefore, negative TCR gamma results in CTCL should be taken with caution. The detection of clonal TCR gamma gene rearrangement by PCR based method is an adjuvant diagnostic marker for CTCL, although it can be seen in some benign dermatoses

Molecular hematology. Qualitative to quantitative techniques

Over the last decade molecular diagnostics technology has developed dramatically from the most laborious, time- consuming southern blot methodology through the revolution of polymerase chain reaction PCR technology to the most reliable, fast, and contamination free molecular analyzer, the real-time quantitative-PCR. The Section of Hematology, Department of Pathology and Laboratory Medicine at King Faisal Specialist Hospital and Research Center has shared this experience during the last 10 years with more than 6,546 samples submitted for the analysis of different gene rearrangements, fusion gene transcripts and gene mutations including Ig heavy chain gene rearrangement for B-cell malignancies, T-cell receptor gamma chain gene rearrangement for T-cell malignancies, BCR/ABL-P210 and P190 fusion gene transcripts, for chronic myeloid leukemia and Philadelphia positive acute lymphoblastic leukemia, PML/RARa fusion gene for promyelocytic leukemia, AML1/ETO for acute myeloid leukemia AML-M2 with t8;21, CBFB/MYH11 for AML M4E0 with inv 16, BCL-2 for follicular lymphoma, and BCL-1 for mantle cell lymphoma. Hence, most molecular assays are qualitative in nature, quantitative assays are deemed necessary in the monitoring and follow-up of minimal residual disease in leukemia and lymphoma, and proved in our experience to serve as an essential tool to confirm complete remission CR post-chemotherapy and bone marrow transplantation, and to detect signs of early relapse for proper clinical intervention. In this manuscript, we retrospectively review our experience in molecular hematology and propose our recommended guidelines at King Faisal Specialist Hospital and Research Center

Reference range values of hematological parameters in saudi healthy adults

This study has been designed and conducted to establish the normal values of various hematological parameters for healthy adult Saudi males and females, and to compare these values with those obtained for other populations in both Western and tropical countries. The study was undertaken in a reference adult Saudi population in the area of Riyadh, Saudi Arabia. A total of 300 [150 males and 150 females] healthy volunteers whose ages ranged between 20-40 years, were investigated. All laboratory analysis was conducted under standardized conditions at the Hematology Section, Department of Pathology, King Faisal Specialist Hospital and Research Center. In males, the mean hemoglobin concentration of 15.2 g/dl [range 13.2-17.20 g/dl], and hematocrit ratio of 0.45 [range 0.39-0.51] were significantly higher than female values of 12.9 g/dl [range 11.0-14.7 g/dl] and 0.38 [range 0.33-0.44] [p<0.05]. The mean red cell count values of 5.2 x 10[12] /L [range 4.4-6.0 x 10[12] /L] in males were also significantly higher than the corresponding values of 4.5 x l0[12] /L [range 3.8-5.3 x l0[12] /L] in females [p<0.05]. The mean and range values of mean cell volume in males [mean of 86.40 fl and range of 82.20-94.90 fl] were significantly higher than in females mean of 84.0 and range of 73.10-94.90 [p<0.05]. Similarly, the mean and range values of mean cell hemoglobin and mean cell hemoglobin concentration were significantly higher in males than the corresponding values in females [p<0.05]. On the other hand, the mean white blood cell count of 6.5 x 10[9] /L [range 3.3-9.7 x 10[9] /L] in the males was significantly lower than the mean value of 7.2 x 10[9] /L [range 3.60-12.80, x 10[9] /L] in the females [p<0.05]. Similarly, the values for platelet counts of 246 x 10[9] /L [range 156-377 x 10[9] /L] in the males were also significantly lower than the corresponding values of 283 x 10[9] /L [range 169-396 x 10[9] /L] in the females [p<0.05]. This study has established base-line values for hematological parameters in healthy Saudi adults. The sex difference of the measured levels of all of these parameters has attained statistical significance. Furthermore, when the established values of this study were compared with those quoted for the methods in use and those drawn from different populations, several differences emerged. Such differences are of importance for the accurate clinical interpretation of hematological investigations of patients

Application of polymerase chain reaction to detect rearrangement of immunoglobulin heavy chain genes in lymphoproliferative disease

As part of our routine work-up in the diagnosis of lymphoproliferative disease, we used a rapid polymerase chain reaction [PCR] assay to amplify the DNA fragments of the framework 3 [FR3] region of the immunoglobulin heavy [IgH] chain genes. The assay does not involve hybridization, nested priming, or sequencing of the amplified PCR product. It was performed on 66 specimens of B-cell lymphoproliferative disease, including acute lymphoblastic leukemia, chronic lymphocytic leukemia, multiple myeloma, hairy cell leukemia and follicular lymphoma. Twenty-six specimens of negative controls, including acute myeloid leukemia, chronic myeloid leukemia in myeloid transformation and idiopathic thrombocytopenic purpura, were also analyzed. The assay was performed with 77% sensitivity and 100% specificity. The standard IgH chain gene rearrangement by Southern blot analysis is reserved for the remaining negative cases if clinically indicated

Detection of BCL-2 gene rearrangement in follicular lymphoma by polymerase chain reaction and chemiluminescence technique

The incidence of follicular lymphoma in Saudi Arabia is very low compared to that in Western countries. We analyzed 22 diagnosed cases, based on conventional morphology examination and immunohistochemistry, to detect the Bcl-2 gene rearrangement by polymerase chain reaction [PCR]. The DNA was extracted from formalin-fixed paraffin-embedded lymph node tissues by the standard xylene treatment and proteinase K digestion method. Rearrangement of the major breakpoint region was evident in 8 of the 22 cases [36%], determined by visualization of a discrete band hybridized with a chemiluminescence-labeled specific probe. Although the number of cases is small, we believe it denotes a normal detection rate for PCR analysis, using DNA isolated from fixed tissue. With the exception of follicular lymphoma, non-Hodgkin's lymphoma [NHL] analyzed included diffuse large cell lymphoma, lymphoblastic lymphoma, chronic lymphocytic leukemia, mucosa-associated lymphoid tissue and mantle zone lymphomas. No Bcl-2 gene rearrangement was detected in any of these cases. No evidence of Bcl-2 minor cluster sequence gene rearrangement was detected in any of the 38 NHL cases analyzed

Immunophenotyping of acute leukemia at King Faisal specialist hospital and research centre

During a 20 month period, 133 bone marrow samples from an equal number of patients with acute leukemia were immunophenotyped. Patients ranged in age from two to 68 years with a mean of 23 years. Eighty-four [63.2%] were classified as acute lymphocytic leukemia [ALL] with the following immunologic subclassification: common ALL 83.3%, T-cell ALL 11.9%, null-cell ALL 2.4% and 2.4% differentiated B-cell ALL. Twenty-eight cases [21%] were classified as acute myeloid leukemia [AML] and 16 cases [12%] demonstrated biphenotypic features. Concordance with morphology and cytochemistry was observed in 129 cases [97%]. Four cases [3%] manifested discrepancy between immunophenotyping, morphology and cytochemistry. We conclude that immunophenotyping by flow cytometry is a useful and reliable method for classification of acute leukemia, especially when interpreted in the light of morphology and cytochemistry

Multiple myeloma: a review of 92 cases at King Faisal Specialist Hospital and Research Centre, Riyadh, Saudi Arabia

A review of 92 cases of multiple myeloma [66 males and 26 females] seen at the King Faisal Specialist Hospital and Research Centre from October 1975 through December 1987 revealed the age for affected patients ranged from 23 to 90 years [mean, 56 years]. Six percent of the patients were less than 40 years old at the time of diagnosis. Bone pain was the most common presenting symptom in our patients [80%], most frequently involving the back. Anemia was the initial finding in 74%, followed by plasmacytoma [22.8%], hypercalcemia [19.6%], and renal insufficicency [18.5%] Skeletal survey abnormalities were seen in 92. 4% of the cases, with osteolytic lesions as the predominant finding. Serum protein electrophoresis showed a monoclonal paraprotein in 78% of the cases, of which 55.5% were the IgG class. Free light chains were detected in the urine of 20 patients. The median survival time for all patients was 68 months. Twenty patients died of renal failure and/or infection. The combination of melphalan and prednisone was used for treatment in 37 patients, while 31 patients received the M2 protocol and 19 patients received different therapy such as VCEP [vindesine, cyclophosphamide, VP 16 and prednisone], MPV [melphalan, prednisone, and vincristine] or high-dose melphalan. Five patients either refused treatment or died before treatment could be started

Monoclinical gammopathy: ten years' experience at King Faisal Specialist Hospital in Riyadh, Saudi Arabia

One hundred eleven cases of monoclonal gammopathy seen between 1979 and 1986 were reviewed. The mean age of the patients was 53 years, and the male to female ratio was 1.5:1. Of 111 cases, 66 [59.5%] had plasma cell dyscrasia, 22 [19.8%] had malignant lymphoma or carcinoma, eight [7.2%] had hematological disease, 12 [10.8%] had inflammatory conditions, and three [2.7%] had other miscellaneous associated conditions of undetermined orign. Of patients with plasma cell dyscrasia, 50 had multiple myeloma, five had light-chain disease, four had Waldenstrom's macroglobulinemia, and seven had solitary plasmacytoma. I[g]G accounts for 78.4% of monoclonal gammopathy followed by I[g]A [9.9%], I[g]M [7.2%], and free light chains [4.5%]. The patients with plasma cell dyscrasia ranged from 33 to 90 years with a mean age of 57 years, and male to female ratio of 2:1. In some conditions, the association of monoclonal gammopathy may be casually related, while in other conditions, the association may be coincidenta

Intestinal parasites: incidence and etiology in over 1,000 patients at King Faisal Specialist Hospital in Riyadh

Stool specimens from 3, 817 patients were examined for presence of intestinal parasites during the calendar year 1985 at the King Faisal Specialist Hospital and Research Centre, Riyadh. Protozoan or metazoan parasites were found in 1, 036 [27.8%] patients, yielding a total of 1, 411 organisms. Incidence of potentially pathogenic parasites was 22.9% of the total and commensal was 77.1%. Of the first group, Giardia lamblia was most common, constituting over half of all the parasites within this group, followed by Hymenolepis nana. Among the commensals, Entamoeba coli was encountered most frequently, followed by Endolimax nana and Blastocystis hominis. Although over 77% of the parasites observed were considered non-pathogenic, their presence in the stool strongly suggests the possibility of patient's exposure to environmental conditions that may result in the ingestion of contaminated food or water containing pathogenic parasites