RESUMEN
A gold nanorods (GNRs) nonaggregation-based colorimetric probe has been developed for the detection of S(2-) based on that the longitudinal surface plasmon resonance absorption wavelength (LPAW) of GNRs red shifts (Δλ) and the color of the solution distinctly changes on account of the faster stripping of GNRs along longitudinal axis than transverse axis in the process of GNRs reacting with S(2-) ions to form Au(2)S complexes on the GNRs surfaces. The GNRs probe exhibits highly sensitive and selective response toward S(2-) with a wide linear range from 10.0 to 10000.0 µM. The proposed colorimetric probe can be used to visibly detect S(2-) in water samples on line in 15 min with the results agreeing well with those of the optical sensor, showing its great practicality. Moreover, the detection mechanism of the probe is also discussed.
Asunto(s)
Oro/química , Nanotubos/química , Sulfuros/análisis , Resonancia por Plasmón de Superficie , Absorción , ColorimetríaRESUMEN
Using Pb(2+) as ion perturber, phenosafranine (PF) and fluorescein isothiocyanate (FITC) could emit strong and stable room temperature phosphorescence (RTP) signal on the filter paper, respectively. When they were mixed, the phenomenon that the RTP signal of PF and FITC enhanced significantly was found. And 1.12 ag DNA spot(-1) (sample volume was 0.40 µL, corresponding concentration was 2.8 × 10(-15) g mL(-1)) could cause the RTP signal of both PF and FITC to enhance sharply. The content of DNA was proportional to the ΔI(p) of PF and FITC in the system at 634 and 659 nm. Thus, a new solid substrate room temperature phosphorimetry (SSRTP) for the determination of trace DNA was established by using FITC-PF as double-luminescent phosphorescence probe. The detection limit (LD) of this method calculated by 3S(b)/k was 14 zg DNA spot(-1) for PF and 18 zg DNA spot(-1) for FITC, respectively, showing high sensitivity. It has been applied to the determination of trace DNA in practical samples and the analysis results were in accordance with those of fluorescence probe. The reaction mechanism of SSRTP for the determination of trace DNA was also discussed.
Asunto(s)
ADN/análisis , Fluoresceína-5-Isotiocianato/química , Fenazinas/química , Análisis Espectral/métodos , Límite de Detección , Luminiscencia , Sondas MolecularesRESUMEN
The 8-quinolineboronic acid phosphorescent molecular switch (abbreviated as PMS-8-QBA. Thereinto, 8-QBA is 8-quinolineboronic acid, and PMS is phosphorescent molecular switch) was found for the first time. PMS-8-QBA, which was in the "off" state, could only emit weak room temperature phosphorescence (RTP) on the acetyl cellulose membrane (ACM). However, PMS-8-QBA turned "on" automatically for its changed structure, causing that the RTP of 8-QBA in the system increased, after PMS-8-QBA-WGA (WGA is wheat germ agglutinin) was formed by reaction between -OH of PMS-8-QBA and -COOH of WGA. More interesting is that the -NH(2) of PMS-8-QBA-WGA could react with the -COOH of alkaline phosphatase (AP) to form the affinity adsorption (AA) product WGA-AP-WGA-8-QBA-PMS (containing -NH-CO- bond), which caused RTP of the system to greatly increase. Thus, affinity adsorption solid substrate room temperature phosphorimetry using PMS-8-QBA as labelling reagent (PMS-8-QBA-AA-SSRTP) for the determination of trace AP was established. The method had many advantages, such as high sensitivity (the detection limit (LD) was 2.5zgspot(-1). For sample volume of 0.40mulspot(-1), corresponding concentration was 6.2x10(-18)gml(-1)), good selectivity (the allowed concentration of coexisting material was higher, when the relative error was +/-5%), high accuracy (applied to detection of AP content in serum samples, the result was coincided with those obtained by enzyme-linked immunoassay), which was suitable for the detection of trace AP content in serum samples and the forecast of human diseases. Meanwhile, the mechanism of PMS-8-QBA-AASSRTP was discussed. The new field of analytical application and clinic diagnosis technique of molecule switch are exploited, based on the phosphorescence characteristic of PMS-8-QBA, the AA reaction between WGA and AP, as well as the relation between AP content and human diseases. The research results promote the development and interpenetrate among molecule switch technique, lectin science and SSRTP.
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Fosfatasa Alcalina/química , Ácidos Borónicos/química , Enfermedad , Mediciones Luminiscentes/métodos , Quinolinas/química , Aglutininas del Germen de Trigo/química , Adsorción , Fosfatasa Alcalina/metabolismo , Humanos , Límite de Detección , Mediciones Luminiscentes/instrumentación , Mediciones Luminiscentes/normas , Estructura Molecular , Sensibilidad y Especificidad , Temperatura , Aglutininas del Germen de Trigo/metabolismoRESUMEN
A novel solid substrate-room temperature phosphorimetry (SS-RTP) was developed for determination of bumetanide (BMTN). It was validated by determining selectivity, linearity, accuracy, precision, and signal to noise ratio (S/N) for analysis. And all the experiments presented in this work were based on that BMTN inhibited the formation of [Fe-morin](3+) ([FeR](3+)) complex by the reaction between Fe(3+) and R, which led to severe quenching of room temperature phosphorescence (RTP) signal. The rate constant of the reaction (k) was 2.44 x 10(-4) s(-1), the activation energy (E) was 21.39 kJ mol(-1). Detection limit of this method (LD, 5.0 ag spot(-1), corresponding concentration was 1.2 x 10(-14) g mL(-1)) was evaluated and compared with other methods, indicating better sensitivity for BMTN determination using this technique. And due to the high sensitivity of the method, it has been successfully applied to determine BMTN in human urine samples. The linear range was from 0.040 pg mL(-1) to 4.0 pg mL(-1), allowing wide determined range of BMTN. Meanwhile, the mechanism of this method was also discussed.
Asunto(s)
Bumetanida/orina , Flavonoides/química , Hierro/química , Mediciones Luminiscentes/métodos , Compuestos Organometálicos/química , Urinálisis/métodos , Bumetanida/química , Humanos , Concentración de Iones de Hidrógeno , Cinética , Modelos Lineales , Masculino , Adulto JovenRESUMEN
8-Quinolineboronic acid phosphorescent molecular switch (8-QBA-PMS) in the "off" state emitted weak room temperature phosphorescence (RTP) of 8-QBA on the acetylcellulose membrane (ACM) with the perturbation of Pb(2+). When 8-QBA-PMS was used to label concanavalin agglutinin (Con A) to form 8-QBA-PMS-Con A based on the reaction between -OH of 8-QBA-PMS and -COOH of Con A, 8-QBA-PMS turned "on" automatically due to its structure change, and RTP of the system increased 2.7 times. Besides, -NH(2) of 8-QBA-PMS-Con A could carry out affinity adsorption (AA) reaction with the -COOH of alpha-fetoprotein variant (AFP-V) to form the product Con A-AFP-V-Con A-8-QBA-PMS containing -NH-CO- bond, causing the RTP of the system to further increase. Moreover, the amount of AFP-V was linear to the DeltaI(p) of the system in the range of 0.012-2.40 (fg spot(-1)). Thus, a new affinity sensitive adsorption solid substrate room temperature phosphorimetry using 8-QBA-PMS as labelling reagent (8-QBA-PMS-AASSRTP) for the determination of AFP-V was proposed with the detection limit (LD) of 9 x 10(-15) g mL(-1). It had been used to determine AFP-V in human serum with the results agreeing with enzyme-link immunoassay (ELISA), showing promise for the prediction of PHC due to the intimate association between AFP-V and primary hepatocellular carcinoma (PHC). The mechanism of the promethod was also discussed.
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Ácidos Borónicos/química , Carcinoma Hepatocelular/química , Neoplasias Hepáticas/química , Sustancias Luminiscentes/química , Quinolinas/química , alfa-Fetoproteínas/análisis , Carcinoma Hepatocelular/sangre , Humanos , Neoplasias Hepáticas/sangre , Mediciones Luminiscentes , Isoformas de Proteínas/sangre , Sensibilidad y Especificidad , TemperaturaRESUMEN
A new phosphorescent labelling reagent consisting of fullerol, fluorescein isothiocyanate and N,N-dimethylaniline (F-ol-(FITC)(n)-DMA) was developed. The mode of action is based on the reactivity of the active -OH group in F-ol with the -COOH group of FITC to form an F-ol-(FITC)(n)-DMA complex containing several FITC molecules. F-ol-(FITC)(n)-DMA increased the number of luminescent molecules in the biological target of WGA-AP-WGA-F-ol-(FITC)(n)-DMA (WGA and AP are wheat germ agglutinin and alkaline phosphatase, respectively) which improved the sensitivity using solid substrate room temperature phosphorimetry (SSRTP) detection. The proposed method provided high sensitivity and strong specificity for WGA-AP. The limit of detection (LD) was 0.15 ag AP spot(-1) for F-ol and 0.097 ag AP spot(-1) for FITC in F-ol-(FITC)(n)-DMA, which was lower than the method using single luminescent molecules of F-ol-DMA and FITC-DMA to label WGA (0.20 ag AP spot(-1) for F-ol-DMA and 0.22 ag AP spot(-1) for FITC-DMA). Results for the determination of AP in human serum were in good agreement with those obtained by enzyme-linked immunosorbent assay. The mechanism of F-ol-(FITC)(n)-DMA labelling of WGA was discussed.
Asunto(s)
Fosfatasa Alcalina/sangre , Compuestos de Anilina/química , Colorantes Fluorescentes/química , Mediciones Luminiscentes , Enfermedad/etiología , Humanos , Indicadores y Reactivos , Isotiocianatos/química , Espectrometría de Fluorescencia , Aglutininas del Germen de Trigo/químicaRESUMEN
OBJECTIVE: To investigate the factors related to delay in pathology reporting and to improve the quality of pathology service. METHODS: A total of 24 months pathology reports (total number = 21,038) issued by Department of Pathology, Dongyang People's Hospital (randomly selected during the period from 1999 to 2006) were analyzed. The timeliness of these reports was studied with respect to the types of specimens (biopsy versus surgical versus frozen section). The causes for delay in reporting were statistically analyzed. RESULTS: Among all the cases studied, 19,579 reports were timely issued (timeliness rate = 93.06%), whereas 1459 reports were delayed (delay rate = 6.94%). Of the 1459 delayed reports, routine biopsy specimens accounted for 6.02% (665/11,052), surgical specimens for 7.26% (643/8858) and frozen section specimens for 13.39% (151/1128). The main causes for delay in reporting were technical (1158 cases or 79.37%), including requests of immunohistochemical staining, intradepartmental or external consultation, patient contact and discussion with requesting clinicians. Factors related to responsibility, such as inadequate clinical information in the pathology request forms, were identified in 301 cases (20.63%). The delay in reporting was mainly due to factors occurring within the pathology department (which accounted for 1048 cases or 71.83% (P < 0.05) and most were technical (1017 cases or 97.04%). Extradepartmental factors, mainly related to responsibility, were noted in 270 cases (65.69%, chi2 = 709.59, P < 0.05). CONCLUSION: Factors related to delay in pathology reporting can be categorized into technical and responsibility. The former can be rectified by improvement of laboratory procedures, while the latter needs education of the personnel concerned.
Asunto(s)
Servicio de Patología en Hospital , Patología Quirúrgica , Biopsia , Control de Formularios y Registros , Secciones por Congelación , Humanos , Control de Calidad , Distribución Aleatoria , Factores de TiempoRESUMEN
Fluorescein (HFin) emitted strong and stable room temperature phosphorescence (RTP) on filter paper after set at 50 degrees C for 10 min using Li(+) as the ion perturber. HFin existed as Fin(-) when the pH value was in the range of 5.45-7.36. Fin(-) could react with [Cu(BPY)(2)](2+) (BPY: alpha,alpha-bipyridyl) to produce ion association complex [Cu(BPY)(2)](2+).[(Fin)(2)](2-), which could enhance the RTP signal of Hfin. In the presence of bovine serum albumin (BSA), the -COOH group of Fin(-) in the [Cu(BPY)(2)](2+).[(Fin)(2)](2-) could react with the -NH(2) group of BSA to form the ion association complex [Cu(BPY)(2)](2+).[(Fin-BSA)(2)](2-), which contained -CO-NH- bond. This complex could sharply enhance the RTP signal of Hfin and the Delta I(p) was directly proportional to the content of BSA. According to the facts above, a new solid substrate-room temperature phosphorimetry (SS-RTP) for the determination of trace protein had been established using the ion association complex [Cu(BPY)(2)](2+).[(Fin)(2)](2-)as a phosphorescent probe. This method had wide linear range (0.40 x 10(-9)-280 x 10(-9)mg l(-1)), high sensitivity (the detection limit (LD) was 1.4 x 10(-10)m gl(-1)), good precision (RSD: 3.4-4.9%) and high selectivity (the allowed concentration of coexistent ions or coexistent materials was high). It had been applied to the determination of the content of protein in 10 kinds of real samples, and the result agreed well with pyrocatechol violet-Mo (VI) method (P.V.M.M.), which indicated it had high accuracy. Meanwhile, reaction mechanism for the determination of trace protein with [Cu(BPY)(2)](2+).[(Fin)(2)](2-) phosphorescent probe was also discussed. The academic thought of this research could not only be used to develop many kinds of ion association complex phosphorescent probes, but also provided a new way to promote the sensitivity of SS-RTP.
Asunto(s)
Fluoresceína/química , Iones/química , Sustancias Luminiscentes/química , Mediciones Luminiscentes/métodos , Albúmina Sérica Bovina/análisis , Animales , Bovinos , Desecación , Modelos Lineales , Mediciones Luminiscentes/instrumentación , Oxígeno/química , Sensibilidad y Especificidad , Albúmina Sérica Bovina/química , Temperatura , Agua/análisisRESUMEN
A new phosphorescent labeling reagent named self-ordered ring (ESOR) of eosin Y (E) was developed. And the application of the determination of bioactive matter by affinity adsorption solid substrate-room temperature phosphorimetry (AA-SS-RTP) based on ESOR labeling lectin was studied. Results showed that pink and homogeneous ESOR could be formed by E on polyamide membrane (PAM) in the presence of cetyltrimethylammonium bromide (CTAB) and ammonia water. ESOR could emit strong and stable room temperature phosphorescence (RTP) signal of E in the presence of heavy atom perturber. Specific affinity adsorption (AA) reactions could be carried out between the products of concanavalin agglutinin (Con A), triticum vulgaris lectin (WGA) labeled with ESOR and alpha-fetoprotein variant (AFP-V), alkaline phosphatase (ALP), glucose (G), respectively. Not only did the products of the affinity adsorption reactions preserve good RTP characteristic of E, but also the DeltaIp (DeltaIp=Ip2-Ip1, Ip1 is the RTP intensity of blank reagent, Ip2 is the RTP intensity of sample) of these products was proportional to the content of AFP-V, ALP and G, respectively. According to the facts above, a new method of AA-SS-RTP for the determination of AFP-V, ALP and G was established, based on ESOR labeling lectin. Detection limits (LD) of this method were 0.040 fg spot(-1) for AFP-V, 0.045 fg spot(-1) for ALP and 0.090 fg spot(-1) for G. And it has been successfully applied to the determination of AFP-V in human serum as well as the survey and forecast of human diseases. This method had high sensitivity, good repeatability, long RTP lifetime and little background interference with lamdamaxem at the long wavelength area. Meanwhile, the mechanism for the determination of trace AFP-V by AA-SS-RTP based on Con A labeled with ESOR was also discussed.
Asunto(s)
Fosfatasa Alcalina/análisis , Eosina Amarillenta-(YS)/química , Glucosa/análisis , Temperatura , Aglutininas del Germen de Trigo/química , alfa-Fetoproteínas/análisis , Adsorción , Concanavalina A/química , Humedad , Mediciones Luminiscentes/instrumentación , Mediciones Luminiscentes/métodos , Estructura Molecular , Oxígeno/química , Sensibilidad y Especificidad , Espectrofotometría/instrumentación , Espectrofotometría/métodos , Coloración y Etiquetado , Factores de TiempoRESUMEN
In this paper, 3.5-generation polyamidoamine dendrimers (3.5G-D)-porphyrin (P) dual luminescence molecule (3.5G-D-P) was developed as a new phosphorescence-labeling reagent. Meanwhile, the room temperature phosphorescence (RTP) characteristics of 3.5G-D-P and its product of labeling triticum vulgaris lectin (WGA) on the surface of polyamide membrane (PAM) were studied. Results showed that in the presence of heavy atom perturber LiAc, 3.5G-D and P of 3.5G-D-P molecule could emit strong and stable RTP on the PAM. And the Tween-80 would spike thoroughly the phosphorescence signal of 3.5G-D and P; moreover, specific affinity absorption (AA) reaction between the products (Tween-80-3.5G-D-P-WGA) of WGA labeled with Tween-80-3.5G-D-P and glucose (G) was carried out. The products of the AA reaction could keep good RTP characteristics of 3.5G-D and P dual luminescence molecule, and the DeltaI(p) was linear correlation to the content of G. According to the facts above, a new method of affinity adsorption solid substrate-room temperature phosphorimetry (AA-SS-RTP) for the determination of trace G was established, basing on WGA labeled with Tween-80-3.5G-D-P dual luminescence molecule. The detection limit of this method was 0.13fgspot(-1) (1.7x10(-12)moll(-1), 3.5G-D) and 0.14fgspot(-1) (2.2x10(-12)moll(-1), P). Determination of G in human serum using excitation/emission wavelength of either 3.5G-D or P, the result was coincided with enzyme-linked immunosorbent assay (ELISA). Not only the sensitivity and accuracy of this method were higher, but also the flexibility of AA-SS-RTP was obviously improved and the applicability was wider.
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Dendrímeros/química , Enfermedad , Glucosa/análisis , Porfirinas/química , Aglutininas del Germen de Trigo/química , Adsorción , Humanos , Luminiscencia , Análisis Espectral/métodos , TemperaturaRESUMEN
In the presence of ion perturber LiAc, 4-generation polyamidoamine dendrimers (4G-D) could emit strong and stable room temperature phosphorescence (RTP) signal at lambda(max)(ex)/lambda(max)(em) = 511.8/675.3 nm on nitrocellulose membrane (NCM), and Triton X-100 could sharply enhance the RTP signal of 4G-D. Triton X-100-4G-D was used to label concanavalin agglutinin (Con A) to get the labeling product Triton X-100-4G-D-Con A. Quantitative specific affinity adsorption (AA) reaction between Triton X-100-4G-D-Con A and alpha-fetoprotein variant (AFP-V) could be carried out on the surface of NCM, whose product Triton X-100-4G-D-Con A-AFP-V could emit strong and stable RTP and its deltaI(p) was proportional to the content of AFP-V. According to the facts above, a new affinity adsorption solid substrate-room temperature phosphorimetry (AA-SS-RTP) for the determination of trace AFP-V by Con A labeled with Triton X-100-4G-D was established. Detection limits of this method were 0.23 fg spot(-1) (direct method, corresponding concentration: 5.8x10(-13) g mL(-1)) and 0.13 fg spot(-1) (sandwich method, corresponding concentration: 3.2x10(-13) g mL(-1)). It has been successfully applied to determine the content of AFP-V in human serum and forecast human diseases, for its high sensitivity, long RTP lifetime, good repeatability, high accuracy and little background perturbation with lambda(max)(em) at the long wavelength area. Meanwhile, the mechanism for the determination of trace AFP-V using AA-SS-RTP was also discussed.
Asunto(s)
Pruebas Diagnósticas de Rutina , Mediciones Luminiscentes/métodos , Suero/química , alfa-Fetoproteínas/análisis , Adsorción , Variación Genética , Humanos , Valor Predictivo de las Pruebas , Sensibilidad y Especificidad , Temperatura , alfa-Fetoproteínas/genéticaRESUMEN
Nitrocellulose membrane-poly (vinyl alcohol)-ionic imprinting (NCM-PVA-I-I) was prepared using Cu2+ as template. The cavity in NCM-PVA-I-I matched Cu2+ very well and the selectivity was high. Cu2+ entered the cavity and then could form ionic association ([Cu2+] x [(Fin-)2]) with the anion of fluorescein (Fin-) outside the cavity by electrostatic effect. [Cu2+] x [(Fin-)2] could emit strong and stable room temperature phosphorescence on NCM-PVA-I-I. Its DeltaI(p) was proportional to the content of Cu2+. Based on the above facts, a new method for the determination of trace copper by solid substrate-room temperature phosphorimetry (NCM-PVA-I-I-SS-RTP, SS-RTP is the abbreviation of solid substrate-room temperature phosphorimetry) using NCM-PVA-I-I technique has been established. The linear range of this method was 2.00-144.00 fg Cu2+ spot(-1) (sample volume: 0.40 microL spot(-1), corresponding concentration: 5.00-360.00 pg mL(-1)), and the detection limit calculated by 3Sb/k was 0.43 fg Cu2+ spot(-1) (corresponding concentration: 1.1 x 10(-12) g mL(-1), n=11). Samples containing 2.00 and 144.00 fg Cu2+spot(-1) were measured, respectively, for seven times and R.S.D.s were 3.5% and 4.7%. NCM-PVA-I-I-SS-RTP could combine very well the characteristics of both the high sensitivity of SS-RTP and the high match and selectivity of NCM-PVA-I-I, and it was rapid, accurate, sensitive and with good repeatability. It has been successfully applied to determine trace copper in human hair and tea samples.
Asunto(s)
Colodión/química , Cobre/análisis , Alcohol Polivinílico/química , Iones , TemperaturaRESUMEN
A new solid substrate-room temperature phosphorescence (SS-RTP) method for the determination of trace manganese (II) has been established. It bases on the fact that fullerol (R) emits strong and stable room temperature phosphorescence (RTP) on filter paper substrate. H2O2 can oxidize R to cause the SS-RTP quenching. But manganese (II) can obstruct H2O2 to oxidize R, and enhance the RTP of R. alpha,alpha'-Bipyridine (Bipy) can sensitize the RTP. After adding Bipy, the DeltaI(p) enhances 7 times than that without Bipy. Under the optimum conditions, the linear dynamic range of this method is 0.016-1.12 pg spot(-1) with a detection limit (L.D.) of 4.6 fg spot(-1) (m(Mn(2+) is the absolute mass of Mn(2+)), and the regression equation of working curve is DeltaI(p)=25.20 + 63.55 m(Mn(2+) (pg spot(-1)), n=6, r=0.9983. For 0.016 and 1.12 pg spot(-1) Mn(2+), RSDS are 4.3 and 4.8%, respectively (n=7). This method has been applied to the determination of trace manganese (II) in actual sample with high sensitivity and good selection. And the reaction mechanism of SS-RTP is discussed.
Asunto(s)
2,2'-Dipiridil/química , Peróxido de Hidrógeno/química , Indicadores y Reactivos/química , Mediciones Luminiscentes , Manganeso/análisis , Oxidantes/química , Sensibilidad y Especificidad , Espectrometría de Fluorescencia , TemperaturaRESUMEN
OBJECTIVE: To develop a protocol for gene rearrangement study in non-Hodgkin's lymphoma (NHL) by PCR-directed gel-scan method and to set up quantitative criteria for IgH gene rearrangement which can be applied in the follow up of lymphoma patients. METHODS: IgH gene rearrangement studies were carried out in 96 cases of B-cell NHL. The detection rate of clonality was evaluated. Sixty-five cases of IgH gene rearranged cases proven by FR3A-directed PCR and PAGE and 8 cases of benign lymphoid tissues (5 cases of reactive lymphoid hyperplasia, 3 cases of chronic tonsillitis), 5 cases of normal peripheral blood mononuclear cells were analyzed by gel-scan method and the proportion of h1/h2 (heights of peak1 and peak2 of gel-scan) was calculated. RESULTS: The detection rate of IgH gene clonality was up to 68% using primer FR3A in the 96 B-cell NHL cases. The detection rate was up to 61% using primer FR2A. With a combination of primers FR3A and FR2A, the detection rate increased to 83%. Gel-scan curve showed that the value of h1/h2 was greater than 3 in all the 65 cases with IgH gene rearranged. In the 8 benign lymphoid tissue cases showed h1/h2 < 1.5, 5 cases with normal peripheral blood mononuclear cells showed a bell-shaped curve. CONCLUSIONS: In the gel-scan curve of gene rearrangement studies in non-Hodgkin's lymphoma samples, the value of h1/h2 greater than 3 represents a true clonal proliferation. The peaks with relative heights less than 1.5 may not be significant and likely represent polyclonal cell population. A value between 1.5 and 3 however requires clinical follow-up. The success rate of rearrangement studies in B-cell NHL can be increased by using a combination of primers FR3A and FR2A.