Prevention is better than Treatment

OSMMR2000 - G-6-PD KIT

Assay in Brief

  1. Take a clean 96-well (preferably U bottom) microplate (elution microplate).

  2. Add 5 microliters of whole blood or one disk cut from a dried blood spot (3/16 inch or 2 X 1/8inch diameter) per well. You can use one microplate for all kind of samples (whole blood and dried blood spots can be measured simultaneously - see note on step 4 below). Remember to add controls (not included in this kit)*

  3. Add 75 microliters of Elution Buffer in each well, mix well the contents of each well and place the plate in a plate shaker (preferably in an incubator at 37 degrees C)

  4. Wait 20 minutes for dried blood spots or 8 minutes for whole blood samples. Hint : if performing an analysis of both dried blood spots and whole blood samples in the same microplate, you can start the elution of the dried blood spots and add the elution buffer in the whole blood samples 8 minutes before the elution stage is over.

  5. While waiting reconstitute one Reagent Vial with distilled water or water for injection. Add 40 ml / Reagent vial if you are using the 4X500 kit format or 8 ml if you are using the 20X100 kit format.

  6. Add 75 microliters of Reagent to each well in a new microplate (reagent microplate).

  7. Slowly warm the microplate with the reagents up to the temperature you will run the experiment (preferably in an incubator at 37 degrees C)

  8. Once the elution stage is over, take both microplates out of the incubator and transfer 15 microliters of each eluted sample to the corresponding well of the reagent microplate (already containing the reagents). Mix well the contents of each well.

  9. Place the reagent microplate in the incubator and wait for 5-7 minutes.

  10. Place the microplate in the microplate reader. The reader should be set to "kinetic mode" at 340 nm. Take 12-15 readings at 1-minute intervals. If your reader is not equipped with a heating device we encourage running the assay in endpoint mode. In this case you may take two readings only (time = 0 and time = 10-15 minutes). In the meantime, the microplate should remain in the incubator. Calculate the ΔOD / min for each sample and the controls. If you are using the endpoint protocol you should substitute the δOD/min with the δOD observed. (see formulae at bottom of this page)

  11. Set the reader to "endpoint mode" and measure the microplate at 405 nm once.

  12. Perform the Hb Normalization using one of the formulas given below.

* We recommend using the Sigma® Chemical Co G-6-PDH controls. Not included in this kit

Notes on this chart:
The chart above shows the results obtained from samples with progressively higher G-6-PD activity using the two different kits, the OSMMR2000 (340 nm - kinetic mode) and OSMMR2000D (550 nm - two point measurement - endpoint mode). Temperature was set to 37 C and the light path was 1 cm. In a microplate reader  the mOD/min obtained were reduced by 50% for both kits (light path approximately 4 mm). Data produced by R&D Diagnostics Ltd.

Endpoint (two-point) mode:

(δODsample340nm)/(δODcontrol340nm)
---------------------------------------------------------- X Control=Sample (U/g Hb)
ODsample405nm / ODcontrol405nm

Kinetic mode

(δODsample340nm/min)/(δODcontrol340nm/min)
---------------------------------------------------------- X Control=Sample (U/g Hb)
ODsample405nm / ODcontrol405nm

INSTRUCTIONS FOR USE (PACKAGE INSERT)

 

R&D DIAGNOSTICStm

G-6-PD

 

 

 

 

A SCREENING KIT FOR DRIED BLOOD SPOTS

This kit is particularly suitable for screening for G-6-pd deficiency in newborns.

WITH Hb NORMALIZATION© PROCEDURE

MANUFACTURER CODE: GRM7 - EDMA CODE: 13.01.09.01 - PRODUCT FILE No. GR/CA01/GRM7/O/127

 

KIT SIZE: 2000 TESTS

 Manufactured by :

R & D  DIAGNOSTICS LTD

41, El. Venizelou str.,

15561 Holargos

Greece

for

  Version XIX, 5 NOVEMBER 2004

 Catalog No : OS-MMR2000 (One Step kit)

  

BY

R&D Diagnostics Limited, Holargos, Greece.

www.rddiagnostics.com
CONTENTS

 

1)   Intended Use

2)   Clinical Background (Summary and Explanation of the Test)

3)   Principle of the Assay

4)   Kit Contents

5)   Reagents / Materials required but not provided

6)   Precautions - Safety and Operating

7)   Sample Collection

8)   Reagent Reconstitution

9)   Assay Procedures

10) Calculation of Results

11) Quality Assurance

12) Expected Values

13) Comparative Protocol Evaluation

14) Performance Characteristics

15) Limitations of Use

16) Summary of Protocols

17) References

1. Intended Use

 

The OSMMr assay is an enzymatic colorimetric method for the quantitative determination of G-6-pd activity in dried blood spots specimens. The test is intended for use as a screening method for red cell glucose-6-phosphate dehydrogenase deficiency in newborns.

2. Clinical Background (Summary and Explanation of the Test)

 

Glucose-6-phosphate dehydrogenase (G-6-PD) is a cytoplasmic enzyme that is distributed in all cells. It catalyses the first step in the hexose monophosphate pathway, producing NADPH (Picture 1). This coenzyme is required as hydrogen donor for reactions of various biochemical pathways as well as for the stability of catalase and the preservation and regeneration of the reduced form of glutathione. Catalase and glutathione are both essential for the detoxification of hydrogen peroxide, therefor the defence of cells against H2O2 ultimately and heavily depends on G-6-PD. The red cells are exquisitely sensitive for oxidative damage and lack other NADPH-producing enzymes. The defence against oxidizing agents, epitomised by H2O2, is mainly realised by glutathione, which converts H2O2 to H2O stoichiometrically via glutathione peroxidase. NADPH is the hydrogen donor for the regeneration of reduced glutathione. An alternative pathway of H2O2 detoxification is via catalase, but this route is regarded ineffective under normal conditions (1), because of the lower affinity of catalase for H2O2 compared to that of glutathione peroxidase. G-6-PD deficiency is the most common known enzymopathy with around 400 million people affected worldwide. The prevalence ranges from 5 to 25% in endemic areas, such as Africa (2,3), the Midle East, Asia (4,5,6,7), the Mediterranean (8,9,10) and Papua New Guinea. The highest incidence is found in Kurdish Jews: 65%. Incidences ranging from 0.5 to 6.9%  have been reported in North and South America. Around 400 mutations have been reported so far (11,12,13,14).

The clinical manifestations associated with G-6-PD deficiency are:

Drug induced hemolysis: certain antimalarials, sulfonamides, sulfones and other drugs or chemicals are associated with significant hemolysis in subjects (15,16,17).

Infection induced hemolysis: numerous bacterial, viral and rickettsial infections have precipitated hemolysis, but the most important are infectious hepatitis, pneumonia and typhoid fever (18,19,20).

Favism: sudden onset of acute hemolytic anemia within 24 to 48 hours of ingesting     fava beans (21,22, 23, 24,25,26,28,28).

Neonatal jaundice: jaundice usually appears by 1 to 4 days of age (29,30,31,32).

Chronic nonspherocytic hemolytic anemia (33).

3. Principle of the Assay

The OSMMr G-6-PD Assay utilizes glucose-6-phosphate-dehydrogenase, which in the presence of NADP, catalyses the oxidation of glucose-6-phosphate  to 6-phosphogluconate.   The NADPH produced is measured colorimetrically at 340 nm in kinetic mode. The results are calculated by evaluating the increase in OD per minute (slope) for unknowns against the slope for a Standard with known G-6-PD activity. Alternatively, the NADPH produced can be measured in endpoint mode using two measurements one at time = 0 and a second one at time = 10-15 minutes later. During the measurements the microplate should remain in the incubator. This protocol is strongly encouraged if your microplate reader is not equipped with a kinetic software or a heating plate. In this case the total OD / sample should be used for calculations. See the attached "Benchtop" instructions for the formulas.

4. Kit Contents

The OSMMr G-6-pd kit contains rubber stoppered glass vials secured with an aluminum crimp seal, which are labeled accordingly. The whole kit is better stored refrigerated. The kit has enough reagents for at least 2000 () tests. The kit comes in two formats: 4 X 50ml reagent vials (each sufficient for at least 500 tests) or 20 X 8ml reagent vials (each sufficient for at least 100 tests).

1 X 200 ml ELUTION AGENT (Code RD6001), Liquid. Contains 0.15% (w/v) sodium azide as preservative. (CAUTION: See safety section 6). Store at room temperature. Refrigeration recommended..

4 X 40 ml or 20 X 8 ml REAGENT VIALS LYOPHILIZED (Code RD6002). Store at 2-8o C (refrigerated – do not freeze) CAUTION: Very toxic to humans.

Stability of the Chemicals

The minimum shelf-life of the chemicals contained in the  kit are as follows :

ELUTION AGENT                                                                               Stable for at least 24 months at room temperature

REAGENT VIAL                                                                                   Stable for 5 days (after reconstitution) at 2-8oC, stable for 5 days

frozen (after reconstitution) plus an additional 5 days following 

thawing. Stable for at least 18 months lyophilized in unopened vial. See Exp.date on sticker.

Although some of the chemicals are stable at room temperature, storing at 2-8oC is highly recommended for all chemicals. This will reduce degradation even more and slow down any microbial infections.

Ongoing stability studies may prove that the chemicals can be stored for much longer periods under certain conditions. This information will be included in the updated temporary insert leaflets and in the final version.

 5. Reagents / Materials required but not provided

1.        U - well micro titration plates (elution plates)

2.        Water for injection (de-ionized water).

3.     Flat-bottomed micro titration plates (assay plates)

4.     Single or multichannel automatic pipettes to deliver volumes in the range of 15 to 75ìl with an accuracy of +/- 1.5% over this range.

5.     A micro titration plate reader capable of reading absorbances at 340 nm  in KINETIC  reading mode.

6.        A hole punch which produces 3/16” or 1/8” diameter discs.

7.        A plate incubator set at 37oC with an accuracy of +/- 1oC

8.     Blood collection cards. The minimum preprinted information required (in accordance to Schleicher and Schuell 2992) is :

1.   Infant’s name, Mother’s name and Patient ID number

2.   Date of Birth

3.   Birth Weight

4.   Sex

5.   Feeding status

6.   Specimen collection date

7.   Submitter’s ID and address

8.   Physician’s name and telephone number

9.   Testing laboratory, name and address

9.             Space for test results

10.           Appropriate number of preprinted ½” (internal diameter) broken or dotted line circles on attached blood collection filter paper.

11.           Manufacturer’s and lot number of filter paper indicated on filter paper

12.           Manufacturer or printer listed on the information section.

                Blood collection filter cards should be attached to at least 2 copies of the information section outlined above.

                Filter paper should can be used according to the specification provided by the National Screening Association.

6. Precautions - Safety and Operating

 All reagents must be stored according to the instructions on the label and instructions for use.

The assay-performance can be seriously affected if the instructions as outlined in this package insert and on the labels are not strictly adhered to.

The Elution Agent contains 0.15% sodium azide as anti microbial preservative. This substance has a toxic effect if absorbed or indigested.  They should  be discarded with an adequate water flow.

All blood samples of human origin must be regarded as a potential biohazard s and normal laboratory precautions must be taken whilst handling these samples.

Do not use kit components after the expiry date stated on the label.

Do not mix components of different lot numbers

Do not use any solutions that have become turbid or discolored.

7. Sample Collection

Collect from the infant's heel according to the NCCLS Approved Standard LA4-A2 "Blood Collection on Filter Paper for neonatal Screening Programs" Second edition (1992) NCCLS, Villanova, PA

After the sample is taken and the blood has dried, the cards must be stored at 2-8oC. Spots not stored under these conditions gradually loose the enzyme activity due to heat inactivation, causing potential risk of misclassifying samples as screen-positive (34,35).

8. Reagent Preparation

The reagents must be warmed up slowly to 30 or 37oC prior to use if these are the temperatures the assay is going to be performed at. No pre-warming needed for temperatures 22-29oC.

Reconstitute 1 VIAL of REAGENT with 40 ml of WATER FOR INJECTION (or de-ionized water). If you are using the 20X100 test format, each reagent vial should be reconstituted with 8 ml of WATER FOR INJECTION (or de-ionized water). Use 4 ml if you are using the 50 test / vial kits or 2 ml if you are using the 25 test / vial kits. The number of tests / vial is shown on the label attached on the reagent vial.

Stable for 5 days at 2-80C or 10  days at -200C. If a frozen aliquot is thawed, this is stable for 4 days at 2-80C

9. Assay Procedure

A) Micro tritration plate:

1) Punch blood spots 3/16 inch diameter (or 2x 1/8" )  in U bottom micro titer plate. Alternatively, you can use 5 microliters of whole blood (with anticoagulant).

    Use position A1, A2 for the Normal Standard, (not provided with the kit).

    Use 2 wells for the Intermediate and Deficient Controls (not provided with the kit.)

2) Add 75 ìl of ELUTION agent to each well.

3) Place micro titer plate on an orbital plate shaker for 30 minutes at room temperature (8 minutes for whole blood samples). This step should be performed at Room Temperature.

4) During the elution, reconstitute and prepare Reagent Mixture. The reagents must be warmed up slowly to 30 or 37oC prior to use if this is the temperature the assay is going to be performed at.

5) Transfer 15ìl of the eluant from each well to the corresponding well of a new flat bottom microtiter plate. If out of range values are obtained the transfer volume can be reduced to 10 microliters.

6) Add 75 ìl of the pre-warmed reagent to each well and mix well.   

7) Place the plate on an orbital shaker for 1 minute.

8) Place the plate in the incubator or reader (if equipped with a heating device) and wait until a total pre-incubation time of 10 minutes (for assay temperature 22-25C) or 7 minutes (assay temperature 25-37C) is completed.

9) Read the plate in a plate reader at 340nm for 15-20 minutes with 60 seconds intervals. Note: if the reaction takes place above room temperature. The reader must be fitted with a temperature-controlled incubator. Alternatively, the NADPH produced can be measured in endpoint mode using two measurements one at time = 0 and a second one at time = 10-15 minutes later. During the measurements the microplate should remain in the incubator. This protocol is strongly encouraged if your microplate reader is not equipped with a kinetic software or a heating plate. In this case the total OD / sample should be used for calculations. See the attached "Benchtop" instructions for the formulas.

10) After the readings are completed, change the program of the reader to an ENDPOINT MODE select wavelength = 405 nm and read the plate (containing the same mixture) once again. See section 10 “OBTAINING RESULTS DIRECTLY EXPRESSED IN U/g Hb”

B)  Cobas Mira   ( 370C )

1) Punch blood spots  2x 1/8 inch in a Sample Cup, leave caps open

2) Add 150 ìl of ELUTION agent to each well.

3) Place Sample Rack on an orbital plate shaker for 30 minutes.

4) During the elution, reconstitute and prepare Reagent Mixture;

    1 test uses 150ìl of Reagent Mixture.

5) Close Sample cups and place Sample Rack on the instrument. Enter the samples in the worklist using the RATIO for G6PD activity and Hb Normalization (see settings attached)

6) Load Reagent Mix for G6PD activity assay and WATER for the Hb Normalization assay.

7) Insert empty cuvettes, and run the test.

See “ Kinetic Measurement of Enzyme Activity – Settings”

C) WHOLE BLOOD SPECIMENS

1) Pipette 10ìl of whole blood into a sample cup

2) Add 150ìl of Elution Agent

3) Vortex mix

Continue at step 4 for Cobas Mira.

D) Hb PROTOCOL Cobas Mira

Run the test using the same sample as used for the G-6-PD activity assay. There is sufficient sample in the cup for a second test. The reagent for the Hb Normalization assay is DILUTION BUFFER.

 “Hemoglobin Evaluation Settings” are available upon request.

10. Calculation of Results

The optical densities for the 15 measurements per sample are plotted against time (in minutes) from which the increase in OD (slopes) for each well can be calculated in OD/min.

These slopes are evaluated against the slope for a Standard with known G-6-PD activity (mean of the slopes of the duplicates). The value for the Standard is divided by slope for the Standard to obtain a factor. This factor is used to multiply the slope's obtained for the unknowns:.

factor = value Control / slope Control

slope unknown x factor = U G-6-PD activity

Example: mean slope Normal Control = 0.015829 OD/min.

               value Control = 17.0 U/mgHb

               factor =   17.0 / 0.015829 = 1074

                slope sample = 0.00845   G-6-PD activity =  9.075 U 

OBTAINING RESULTS DIRECTLY EXPRESSED IN U / g Hb.

The following formula expresses your results directly into U / g Hb (Greek Patent # 1003227; International Patent Pending)

   (dOdsample340nm/min)/(dOdcontorl340nm/min)

                                                                    x Control Value = Sample Value (Activity in U/g Hb)

   Odsample405nm/ODcontrol405nm

wherein dODsample is the change in optical density (per minute) for the sample, 340nm is the wavelength at which optical density is measured under kinetic mode, dODcontrol is the change in optical density (per minute) of a control measured under kinetic mode conditions at a particular wavelength (340nm), ODsample is the optical density for the sample measured once at a particular wavelength (405nm), and ODcontrol is the optical density of the control measured once at that particular wavelength (405nm), 405nm is the wavelength at which a reading of Heamoglobin contained can be taken, the control value being determined by the change in optical density (per minute) of the control measured under kinetic mode at 340nm divided by the optical density of the control at 405nm i.e. dODcontrol340nm/mim/ODcontrol405nm.

EXAMPLES

The Kit described above was used to demonstrate the effectiveness of the second reading on a Cobas Mira Plus ( Roche Diagnostics ).  The experiment was set up to mimic the effect of incomplete elution on the measurement of enzyme activity. According to the assay protocol described above 2 spots of 3.2mm must be punched from the sample and inserted into the sample cup. This was done using  whole blood samples of 3 human donors spotted and dried onto filtration paper. The instrument was programmed for the second reading by repeating the assay from the same sample, only with a single reading at 405nm instead of the kinetic reading at 340nm.  To mimic the effect of incomplete elution the same samples were used but only one spot was punched and inserted into the sample cup.

The results are shown below in Table 1:

Table 1

 

 

mOD/min 340nm

 

 

 

405nm

 

 

Units

 

2 spot

1 spot

%

 

1 spot

2 spot

%

 

 

 

 

 

 

 

 

 

MMRC02

N

19.42

9.98

51.4%

 

1.81

3.28

55.3%

MMRC02

N

23.18

11.2

48.3%

 

1.92

3.81

50.5%

USA

1

8.29

4.2

50.7%

 

1.04

2.07

50.2%

USA

1

7.76

4.71

60.7%

 

1.12

1.97

56.8%

USA

2

12.98

5.98

46.1%

 

1.16

2.48

46.9%

USA

2

13.05

6.07

46.5%

 

1.14

2.40

47.7%

 

 

 

 

 

 

 

 

 

 

 

Ratio

 

 

 

 

Normalised

 

 

 

2 spot

1 spot

 

 

1 spot

2 spot

%

 

 

 

 

 

 

 

 

 

MMRC02

N

1.000

1.810

 

 

18.06

19.42

93.0%

MMRC02

N

1.162

1.705

 

 

19.10

19.96

95.7%

USA

1

0.631

3.154

 

 

13.25

13.14

100.8%

USA

1

0.601

2.932

 

 

13.81

12.92

106.9%

USA

2

0.756

2.823

 

 

16.88

17.17

98.3%

USA

2

0.732

2.866

 

 

17.40

17.84

97.6%

                   

11. Quality Assurance

12. Expected Values

Values Obtained

Cycle No.

Deficient *

Intermediate *

Normal *

5

0

0

0

6

0.0060

-0.0008

0.0067

7

0.0011

0.0009

0.0130

8

0.0001

0.0019

0.0200

9

0.0019

0.0025

0.0273

10

0.0015

0.0035

0.0335

11

0.0029

0.0052

0.0420

12

0.0022

0.0062

0.0481

13

0.0024

0.0068

0.0551

14

0.0030

0.0075

0.0615

15

0.0029

0.0081

0.0667

16

0.0030

0.0094

0.0740

17

0.0033

0.0102

0.0812

·       Values are expressed in ÄODs 

13. Comparative Protocol Evaluation 

14. Performance Characteristics

See attached Quality Control Certificate issued for this batch.

15. Limitations of Use

Low results are not diagnostic per se of G-6-pd deficiency but indicate the need for further study of the newborn from which a presumptive screen positive sample was received.

Important note: Do not perform the assay at temperatures over 37oC as this may inactivate the coenzyme and lead to erroneous results.

In case of bubble formation in the well a FALSE NEGATIVE result may occur, caused by diffusion of the light passing vertically through the well leading to falsely high absorption of light. As a consequence, a G-6-PD Deficient specimen could be misclassified as NORMAL.

16. Summary of Protocol

SEE ATTACHED BRIEF INSERT

17. References

References in the Text

1.Cohen G. and Hochstein P. Generation of hydrogen peroxide in erythrocytes by  haemolytic agents. Biochemistry, 3:895, 1964.

2. Capps F.P.A., Gilles H.M., Jolly H. and Worlledge S.M. Glucose-6-phosphate dehydrogenase deficiency and neonatal jaundice in Nigeria. Their relation to the  prophylactic use of vitamin K. Lancet, ii:379, 1963.

3. Bienzle U., Effiong C.E. and Luzzatto L. Erythrocyte glucose-6-phosphate dehydrogenase deficiency (G6PD type A-) and neonatal jaundice. Acta Paed Scand  65:701, 1976.

4. Lai H.C. Lai M.P.Y. and Leung K.S. Glucose-6-phosphate dehydrogenase deificiency in Chinese. J Clin Pathol 21:44, 1968.

5. Lu T-C, Wei H. and Blackwell R.Q. Increased incidence of severe hyperbilinurinaemia among newborn Chinese infants with G6PD deficiency.  Pediatrics 37:994, 1966

6. Flatz G., Sringam S., Premyothin C., Penbharkkul S., Ketusingh R. and Chulajata R. Glucose 6-phosphate dehydrogenase deficiency and neonatal jaundice. Arch Dis Child  38:566, 1963.

7. Phornphutkul C., Whitaker J.A. and Woramthumrong N. Severe hyperbilinurinemia in Thai newborns in association with erythrocyte G6PD deficiency. Clin Pediatr 8:275, 1969.

8. Doxiades S.A. and Valaes F. The clinical picture of Glucose 6-phosphate dehydrogenase      deficiency in early infancy. Arch Dis Child 39:545, 1964.

9. Doxiades S.A., Fessas P.H. and Valaes F. Erythrocyte enzyme deficiency in unexplained      kernicterus. Lancet ii:44, 1960.

10.Meloni T., Cagnazzo G., Dore A. and Cutillo S. Phenobarbital for prevention of       hyperbilinurinemia in Glucose 6-phosphate dehydrogenase deficient newborn infants. J Pediatr 82:1048, 1973.

11.Matthay K.K and Mentzer W.C. Erythrocyte enzymopathies in the newborn. Clin Haematol 10:31, 1981.

12.Malluh A.A., Imseeh G., Abu-Osba Y.K.and Hamdan J.A. Screening for Glucose 6-phosphate dehydrogenase deficiency can prevent severe neonatal jaundice. Ann Trop  Paed 12:391, 1992.

13.Solem E. Glucose 6-phosphate dehydrogenase deficiency : An easy and sensitive quantitative assay for the detection of female heterozygotes in red blood cells. Clin Chim Acta 142:153, 1984.

14.Solem E., Pirzer C., Siege M., Kollman F., Romero - Savaria O., Barktsch - Trefs O.  and Kornhuber B. Mass screening for Glucose 6-phosphate dehydrogenase deficiency. Improved fluorescent spot test. Clin Chim Acta 152:135, 1985.

15.Dern R.J., Beutler E. and Alving A.S. The haemolytic affect of primaquine. II. The  natural course of the haemolytic anaemia and the mechanisms of its self-limiting character. J Lab Clin Med 44:171, 1954.

16.Beutler E., Dern R.J. and Alving A.S. The haemolytic effect of primaquine. IV. The relationship of cell age to hemolysis. J Lab Clin Med 44:439, 1954.

17.Beutler E. The hemolytic effect of primaquine and related compounds. Blood, 14:103, 1959.

18.Chan T.K., Chesterman C.N., McFadzean A.J.S. and Todd D. The survival of Glucose 6-phosphate dehydrogenase deficient erythrocytes in patients with typhoid fever on chloramphenicol therapy. J Lab Clin Med 77:177, 1971.

19.Burka E.R., Weaver Z. and Marks P.A. Clinical spectrum of hemolytic anemia associated with Glucose 6-phosphate dehydrogenase deficiency. Ann Intern Med  64:817, 1966.

20.Owusu S.K., Addy J., Foli A.K., Janosi M., Konotey - Ahulu F.I.D. and Larbi E.B. Acute reversible renal failure associated with Glucose 6-phosphate dehydrogenase  deficiency. Lancet 1:1255, 1972.

21. Fermi C. and Martinetti P. Studio sul favismo. Ann.Igiene Sper, 15:76, 1905

22.Luisada L. Favism: A singular disease affecting chiefly red blood cells. Medicine,  20:229, 1941

23.Sansone G., Piga A.M. and Segni G. Il Favismo, Torino, Italy, Minerva Medica, 1958.

24.Kattamis C.A., Kyriazakou M. and Chaidas S. Favism : Clinical and biochemical data. J Med Genet 6:34, 1969.

25.Kahn A., Marie J., Desbois J.C. and Boivin P. Favism in a Portuguese family due to a deficient Glucose 6-phosphate dehydrogenase variant of Canton or Canton-like type 1. Acta Haematol 56:58, 1976.

26. Belsey M.A. The epidemiology of favism. Bull WHO 48:1, 1973.

27.Szeinberg A., Sheba C., Hirschorn N. and Bodonyi E. Studies on erythrocytes in cases with past history of favism and drug induced avute hemolytic anemia. Blood 12:603, 1957.

28.Gross A.T., Hurwitz R.A. and Marks P.A. An hereditary enzymatic defect in erythrocyte metabolism. Glucose 6-phosphate dehydrogenase deficiency. J Clin Invest  37:1176, 1958.

29.Matthay K.K. and Mentzer W.C. Erythrocyte enzymopathies in the newborn. Clin Haematol 10:31, 1981.

30.Smith G.D. and Vella F. Erythrocyte enzyme deficiency in unexplained kernicterus. Lancet 1:1133, 1960.

31.Bienzle U. Glucose 6-phosphate dehydrogenase deficiency. Part I : Tropical Africa. Clin Haematol 10:785, 1981.

32.Karayalcin C., Acs H. and Lanzkowsky P. G6PD deficiency and hyperbilinurinemia in black American full-term infants. NY State J Med 79:22, 1979.

33.Luzzatto L. Inherited haemolytic states : Glucose 6-phosphate dehydrogenase deficiency. Clin Hematol 4:83, 1975.

34.Motulsky A.G., Campbell - Kraut J.M. Population genetics of Glucose 6-phosphate       dehydrogenase deficiency of the red cell in : Blumberg B.S. (ed): Proceedings of Conference on Genetic Polymorphisms and Geographic Variations in Disease. New  York: Grune & Stratton, 1961, p.159.

35.Bernstein R.E. Brillinat cresyl blue screening test for demonstrating Glucose 6-phosphate dehydrogenase deficiency in red cells. Clin Chim Acta 8:158, 1963.

36. Reclos G.J., Hatzidakis C.J. and Kruithof R.A. G-6-PD Diagnosis : Modification of the standard method eliminates the need for an additional Hemoglobin determination. Pharmakeftiki 12(1): 25-31, 1999

37. Reclos G.J., Hatzidakis C.J. and Schulpis K.H. Glucose-6-phosphate dehydrogenase deficiency neonatal screening: preliminary evidence that a high percentage of partially deficient female neonates are missed during routine screening. J Med Screening, 7(1): 46-51, 2000

General References

[1] Beutler E. Study of Glucose-6-Phosphate dehydrogenase : History and Molecular Biology. Am.J.Hematol., 42:53, 1993.

[2] Lucio Luzzatto and Atui Mehta, “Glucose 6-Phosphate Dehydrogenase Deficiency, pp. 3367-3398,  in The Metabolic and Molecular Bases of Inherited Disease, by Scriver C.R. et al. McGraw and Hill Editions, 1994.

Contact Information

For Technical Details, Article reprints, Lists of Contraindicated Drugs and other Assay Information you may contact us by

Mail: Send to R&D DIAGNOSTICS LTD, 33, ALEVIZATOU STREET, P.O.BOX 66019, GR 15669 HOLARGOS, GREECE.

Phone: Dial  +30-210-6537307

Fax: Send at +30-210-6548284

E-mail  info@rddiagnostics.com

You may find more information on this Disease and a step by step example of the correct use of the Normalization Procedure in our site at

www.rddiagnostics.com

 

 

About us   The Disorders   The Products   Kit Inserts   Literature   Technical Support   Articles   Events    Links   Contact us   To Order

Hosted by ohmywebsite.com
TO SEARCH THIS SITE - CLICK SEARCH 

Page last edited on 21/03/2006