Question

Case Study #8 A 54-year-old woman with thrombosis, a high platelet count and a decrease erythrocyte sedimentation rate was tested for polycythemia vera. (Polycythemia vera is a slow-growing blood cancer in which your bone marrow makes too many red blood cells. These excess cells thicken your blood, slowing its flow. They also cause complications, such as blood clots, which can lead to a heart attack or stroke.) In the CBC, WBC and neutrophil counts were normal. Iron stores were also within normal range. A blood sample was submitted to the molecular pathology laboratory for JAK2 V617F mutation analysis by multiplex sequence-specific PCR. This mutation is thought to be the cause of Polycythemia vera. The results are shown below: 1 2 3 4 5 Copyright©2012 F.A. Davis Company www.tedavis.com Agarose gel electrophoresis of SSP-PCR amplicons. Extension of a JAK2 V617F-specific primer produces a 270-bpband in addition to the normal 500 and 230-bp bands. Lane 1- Molecular weight markers; lane 2-patient specimen; lane 3-V617F mutant control; lane 4 - normal control; lane 5 -- reagent blank. Questions: 1. What is the source of the two bands in the normal control in lane 4? 2. What is the source of the additional band in the positive control in lane 3? 3. Does this patient have the JAK2 V617F mutation?

          Case Study #8
A 54-year-old woman with thrombosis, a high platelet count and a decrease erythrocyte sedimentation
rate was tested for polycythemia vera. (Polycythemia vera is a slow-growing blood cancer in which your
bone marrow makes too many red blood cells. These excess cells thicken your blood, slowing its flow.
They also cause complications, such as blood clots, which can lead to a heart attack or stroke.) In the
CBC, WBC and neutrophil counts were normal. Iron stores were also within normal range. A blood
sample was submitted to the molecular pathology laboratory for JAK2 V617F mutation analysis by
multiplex sequence-specific PCR. This mutation is thought to be the cause of Polycythemia vera. The
results are shown below:
1
2
3
4
5
Copyright©2012 F.A. Davis Company www.tedavis.com
Agarose gel electrophoresis of SSP-PCR amplicons. Extension of a JAK2 V617F-specific primer
produces a 270-bpband in addition to the normal 500 and 230-bp bands. Lane 1- Molecular weight
markers; lane 2-patient specimen; lane 3-V617F mutant control; lane 4 - normal control; lane 5 --
reagent blank.
Questions:
1. What is the source of the two bands in the normal control in lane 4?
2. What is the source of the additional band in the positive control in lane 3?
3. Does this patient have the JAK2 V617F mutation?

Case Study #8
A 54-year-old woman with thrombosis, a high platelet count and a decrease erythrocyte sedimentation
rate was tested for polycythemia vera. (Polycythemia vera is a slow-growing blood cancer in which your
bone marrow makes too many red blood cells. These excess cells thicken your blood, slowing its flow.
They also cause complications, such as blood clots, which can lead to a heart attack or stroke.) In the
CBC, WBC and neutrophil counts were normal. Iron stores were also within normal range. A blood
sample was submitted to the molecular pathology laboratory for JAK2 V617F mutation analysis by
multiplex sequence-specific PCR. This mutation is thought to be the cause of Polycythemia vera. The
results are shown below:
1
2
3
4
5
Copyright©2012 F.A. Davis Company www.tedavis.com
Agarose gel electrophoresis of SSP-PCR amplicons. Extension of a JAK2 V617F-specific primer
produces a 270-bpband in addition to the normal 500 and 230-bp bands. Lane 1- Molecular weight
markers; lane 2-patient specimen; lane 3-V617F mutant control; lane 4 - normal control; lane 5 –
reagent blank.
Questions:
1. What is the source of the two bands in the normal control in lane 4?
2. What is the source of the additional band in the positive control in lane 3?
3. Does this patient have the JAK2 V617F mutation?

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