Anti-Platelet Glycoprotein Antibodies in Thrombocytosis Are Frequently Detected and Are Associated with Platelet Activation and Thrombosis.

Abstract Introduction: Thrombosis is the major cause of morbidity and mortality in patients with thrombocytosis (TS) associated with myeloproliferative disorders (MPD), but predicting which patients are at risk has been challenging. We recently reported that antiphospholipid antibodies (APLA) and platelet activation are risk factors in TS [C Bidot et al; Hematology, 10:451, 2005]. To shed further light on the cause of platelet activation in TS, we here report on measuring anti-platelet antibodies in a group of these patients. Material and Methods: Thirty-three TS patients, 15 with clinical thrombosis and 18 without, were evaluated using PAICA method of L. Macchi et al [Thromb Haemost76:1020,1996]. We assayed anti-platelet antibodies against 3 target antigens, GPIIb/IIIa (CD41b), GP Ib/IX (CD42b) and GP IV (Cd36) for IgG and IgM. We also measured platelet activation marker CD62P, and microparticles from platelets (PMP, defined by CD31+/CD42+) and endothelial cells (EMP, defined by CD31+/CD42−) by flow cytometry. Results: At least one anti-platelet antibody was detected in 19/33 (57.5%) of TS patients. Considering first IgG in the19 who were anti-platelet antibodies positive, 14/19 (73.6%) were positive for CD36, 12/19 (63%) for CD41b and 8/19 (42.1%) for CD42b. For IgM in these 19 patients, incidences were 58.8% for CD41b, 38.8% for CD36 and 35% for CD42b. When we compared the anti-platelet antibody positive group with the negative group, we observed that activation marker CD62P was significantly higher in the former (p= 0.04), as were EMP levels (p= 0.02), but PMP levels were not different. Next, we compared results with incidence of thrombosis. None of the IgG results discriminated thrombosis from non-thrombosis. However, all three IgM results were significant in this regard: CD36 (p< 0.05), CD42b (p< 0.02), and most notably, CD41b (p< 0.0003). Conclusions: These results show that anti-platelet antibodies of IgM class, especially anti-CD41b, are associated with platelet activation, endothelial activation, and clinical thrombosis in TS. These findings parallel our previous report on APLA in TS. We suggest that IgM-CD41b may activate platelets, predisposing to hypercoagulable state in this disorder.

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Antibodies Against Platelet Glycoprotein Target Antigens in Antiphospholipid Syndrome (APS).

Blood ◽

10.1182/blood.v108.11.3973.3973 ◽

2006 ◽

Vol 108 (11) ◽

pp. 3973-3973 ◽

Cited By ~ 1

Author(s):

Carlos J. Bidot ◽

Wenche Jy ◽

Carlos Bidot ◽

Lawrence L. Horstman ◽

Vincenzo Fontana ◽

...

Keyword(s):

Platelet Activation ◽

Antiphospholipid Syndrome ◽

Antiphospholipid Antibodies ◽

Dominant Role ◽

Endothelial Activation ◽

Platelet Glycoprotein ◽

Activation Markers ◽

Positive Group ◽

Platelet Antibodies ◽

Platelet Microparticles

Abstract Introduction: Antiphospholipid syndrome (APS) is characterized clinically by thrombotic events and the presence of antiphospholipid antibodies (aPLA) and/or lupus anticoagulant (LA). It is frequently associated with thrombocytopenia and anti-platelet antibodies have been implicated by some. However the roles of anti-platelet antibodies in APS have not been elucidated. We previously reported that platelet activation, but not endothelial activation, was associated with thrombosis in aPLA+ patients [Blood, 104:143a, 2004] but the cause of platelet activation was not addressed. In the present study, we investigated the prevalence of anti-platelet antibodies in APS patients, as well as platelet and endothelial activation. Material and Methods: We evaluated 47 patients with primary APS. Anti-platelet antibodies against GP IIb/IIIa (CD41b), GP Ib/IX (CD 42b) and GP IV (CD36) for IgG and IgM class were measured by PAICA assay [Thromb Haemost76:1820, 1996]. We also measured platelet and endothelial activation markers by flow cytometry: CD62P on platelets, CD31+/CD42+ platelet microparticles (PMP), and CD31+/CD42- endothelial microparticles (EMP). Results: Of the 47 patients, 34 (72%) were positive for at least one anti-platelet antibody. Looking first at IgG, 18/34 (53%) were positive for GP IV; 17/34 (50%) for GP IIb/IIIa; and 16/34 (47%) for GP Ib/IX. IgM antibodies were 47% (14/30) for GP Ib/IX, 38%(13/34) for GP IIb/IIIa, and 24% (8/33) for GP IV. Platelet and endothelial markers were significantly more common in the anti-platelet antibodies positive group: 40% vs. 21% for CD62P, 40% vs. 28.5% for EMP, and 23% vs. 5% for PMP, respectively. We found that CD62P associated significantly with IgM anti-GP IIb/IIIa (p< 0.05), and PMP with IgM anti-GP IIb/IIIa (p< 0.05), and IgM anti-GP IV (p< 0.05). Conclusions: Anti-platelet antibodies are common in APS, confirming previous reports. We found that anti-platelet antibodies IgM anti-GP IIb/IIIa, and IgM anti-GP IV were often associated with platelet activation, suggesting that these antibodies may activate platelets to play an important role in the thrombogenesis of APS. These antibodies were also associated with endothelial activation. It remains to be determined which antibodies, APLA and/or anti-platelet antibodies, play a dominant role in the activation of platelet or endothelial cells and contibute most to the pathogenesis of thrombosis in APS.

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Myeloproliferative Disorders Are Associated with an Increase in Levels of Prothrombotic and Inflammatory Markers Despite Cytoreductive Therapy.

Blood ◽

10.1182/blood.v108.11.3625.3625 ◽

2006 ◽

Vol 108 (11) ◽

pp. 3625-3625

Author(s):

Betty Y.Y. Cheung ◽

Beverley J. Hunt ◽

Kiran Parmar ◽

Yvonne Daniel ◽

Deepti Radia ◽

...

Keyword(s):

Platelet Activation ◽

Inflammatory Markers ◽

Disease Duration ◽

Annexin V ◽

Jak2 V617f ◽

Adenosine Diphosphate ◽

Myeloproliferative Disorders ◽

Endothelial Activation ◽

P Value ◽

Polycythaemia Vera

Abstract Thrombotic complications are frequent in patients with Essential Thrombocythaemia (ET) and Polycythaemia Vera (PV). A raised haematocrit and thrombocytosis have been implicated in the pathogenesis but events still occur on treatment. We hypothesized that there was a role for other prothrombotic and inflammatory pathways in the aetiology of thrombosis in MPD, and so assessed these parameters in 101 patients and 51 controls. Assays and results are listed in table below. Standard methods of thromboelastography, flow cytometry, RVVT assays and ELISAs were used. 72 ET and 29 PV patients were included: median age 58 years (range17–89), disease duration 84 months (range 2–384) and platelet counts 473 × 109/l (range 207–1780). 59 (58%) patients had the JAK2 V617F mutation. 51 (50%) had thrombosis with 23 patients developing the events post-diagnosis. 82 (81%) patients were receiving anti-platelet agents and 76 (75%) cytoreductive therapy. Comparison with clinical features revealed V617F positive patients to be older with significantly higher haemoglobin and PCV and greater levels of E-selectin and C3a, contrary to the higher platelet activation recently shown by Arellano-Rodrigo (2006). The only parameters that correlated with previous thrombosis were older age, longer disease duration and increased CD63 levels +/− ADP stimulation. Cytoreductive therapy was associated with lower levels of VEGF (p=0.01) but no other parameters. In conclusion, assessment of haemostatic, endothelial, complement, angiogenic and inflammatory markers shows evidence of increased activation in patients with MPD. Of these parameters, only the angiogenic marker was reduced by cytoreductive therapy and none appear to be associated with the risk of thrombosis. Median values of prothrombotic and inflammatory parameters Parameters Test Units Patients Controls p-value NS=non-significant, PMP=platelet-microparticles, ADP=adenosine diphosphate, PMA= platelet-monocyte aggregates, PGA=platelet-granulocyte aggregates Thromboelastograph R-time secs NS K-time secs 137.5 160 0.002 α- angle degrees 58.7 52.8 <0.0001 MA mm 64.1 59 <0.0001 Functional PMP 0.2μm-filterd plasma 2.35 2.6 <0.0001 0.1μm-filtered plasma 2.42 2.8 <0.0001 Quantitative PMP PMP % 6.41 4.58 <0.0001 Annexin V+ PMP % 0.19 0.15 0.002 Platelet activation Annexin V+ plts % 2.63 2.07 0.004 CD62p % 1.00 0.52 <0.0001 CD62p +ADP % NS CD63 % 3.32 2.12 0.01 CD63 +ADP % 22.2 16.1 0.004 soluble p-selectin mg/ml 172.5 74.7 <0.0001 Platelet-leucocyte aggregates PMA % 12.20 5.94 <0.0001 PGA % 4.56 3.77 <0.0001 Monocyte-tissue factor 0hr % 0.6 0.24 <0.0001 1hr % 1.32 0.62 <0.0001 4hr % NS Endothelial activation soluble ICAM-1 ng/ml 235.9 194.3 0.013 soluble E-selectin ng/ml NS Complement activation plasma C3a ng/ml 113.4 94.2 0.001 Angiogenesis soluble VEGF pg/ml 670 196.2 <0.0001 Inflammatory markers serum IL-8 pg/ml 12.9 11.1 <0.0001 serum CRP, IL-6 NS Haemostasis markers plasma PF 1+2 nmol/l 0.7 0.6 0.027 plasma PAI-1 ng/ml 27.2 21.1 0.001 D-Dimers, tPA, TAFI NS

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Antiphospholipid Antibodies, Platelet and Endothelial Activation in Acute Thrombotic Thrombocytopenic Purpura (TTP).

Blood ◽

10.1182/blood.v104.11.847.847 ◽

2004 ◽

Vol 104 (11) ◽

pp. 847-847

Author(s):

Carlos J. Bidot ◽

Wenche Jy ◽

Joaquin J. Jimenez ◽

Lawrence Horstman ◽

Eugene Ahn ◽

...

Keyword(s):

Platelet Activation ◽

Lupus Erythematosus ◽

Antiphospholipid Antibodies ◽

Endothelial Activation ◽

Blood Chemistry ◽

Igg Antibody ◽

Clinical Recurrence ◽

Platelet Counts ◽

Clinical Observations ◽

Duration Of Hospitalization

Abstract BACKGROUND. Although the etiology of TTP remains controversial, some notable associated conditions have been identified. These include genetic predisposition, infections, drugs, pregnancy, systemic lupus erythematosus (SLE) and deficiency of ADAMTS13. These associations underscore the suspected immunological nature of this disorder. Some authors have reported the presence of antiphospholipid antibodies (APLA) with onset of fulminate severe TTP. Also, platelet and endothelial activation has been described in TTP but their possible relation with APLA is unknown. METHODS. We measured APLA in 19 patients with acute TTP and 17 in remission. Fifteen were female and four were male, mean age of 37 ± 15. All met the diagnostic criteria of TTP and were treated with standard therapy including exchange plasmapheresis. The severity of TTP was classified based on duration of hospitalization and numbers of plasmapheresis needed. Recurrence of TTP was assessed by history and review of medical records. CBC, platelet counts, blood chemistry and LDH were monitored. We investigated APLA antibodies IgG and IgM against 6 antigens (β2GP1, FVII/VIIa, CL, PC, PS, PE) by ELISA. To investigate platelet activation, we measured by flow cytometry: CD62P on platelets, and platelet microparticles (PMP) by CD41 (PMP41). We also measured endothelial microparticles (EMP) using CD62E. RESULTS. Lab study : In acute TTP, 42% (8/19) were APLA positive for at least one IgG antibody. The most frequent was aPS (33%) followed by aCL(28%), aPC(28%), aβ2GP1 (28%), FVII/VIIa (21%) and PE (21%). IgM antibodies were not detected. APLA was two-fold more prevalent in acute TTP (42%) than in remission (21%). In acute TTP multiple antibodies (two or more) were seen in 75% (6/8) while in remission only one (12.5%) had multiple antibodies. Platelet activation measured by CD62P was higher in APLA+ than in APLA− (102 ± 25 vs 33 ± 8 fluorescence units, p<0.02, X ± SEM). PMP41 was higher in APLA+ than in APLA− but this difference was not significant. We also found that EMP62E was significantly higher in APLA+ than in APLA− patients (p<0.01). Clinical observations : When comparing the two groups (APLA+ vs APLA−), APLA+ group had higher incidence of recurrence of TTP than APLA− (75% vs 36%), p<0.05. Platelet counts, LDH, severity and incidence of CNS syndrome were not significantly different between APLA+ and APLA− groups. CONCLUSIONS: APLA are frequently seen in acute TTP (42%) and they tend to decline in remission. Their presence is associated with increased activation of platelets and endothelial cells, and also with clinical recurrence of TTP. APLA may play a role in excerbation and recurrence of TTP.

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Analysis of JAK2V617F Mutation, PRV-1 Overexpression and Endogenous Erythroid Colony (EEC) Formation Show Different Coexpression Patterns among Ph-Negative Chronic Myeloproliferative Disorders.

Blood ◽

10.1182/blood.v106.11.2592.2592 ◽

2005 ◽

Vol 106 (11) ◽

pp. 2592-2592

Author(s):

Beatriz Bellosillo ◽

Carles Besses ◽

Lourdes Florensa ◽

Raquel Longaron ◽

Gemma Navarro ◽

...

Keyword(s):

Direct Sequencing ◽

Specific Treatment ◽

Myeloproliferative Disorders ◽

Jak2v617f Mutation ◽

Negative Group ◽

Jak2 Mutation ◽

Positive Group ◽

Significant Difference ◽

Group A ◽

Group B

Abstract Introduction. Recently, the JAK2V617F mutation has been reported in the majority of PV patients as well as in a variable percentage of ET and IMF patients. Some authors have reported a high correlation of JAK2 mutation with PRV-1 overexpression and the formation of EECs. In the current study we have analyzed the pattern of positivity of these three biomarkers in a cohort of Ph-negative MPD patients. Patients and methods. 103 Ph-negative MPD patients (58 ET, 37 PV and 8 IMF) from a single institution were studied. Patients were diagnosed according to the PVSG and Barosi criteria. EEC formation was determined at diagnosis. At the time when PRV-1 and JAK2 mutation were analyzed 29/103 patients were receiving platelet-lowering therapy ± ASA, 26/103 patients only received ASA and 48/103 received no specific treatment. PRV-1 expression was quantified by real-time reverse transcriptase (RT)-PCR in RNA from granulocytes. Analysis of JAK2V617F was performed by direct sequencing using granulocyte RNA. Results. JAK2 mutation was observed in 21/58 ET (36.2%), 30/37 PV (81.1%) and 5/8 IMF (62.5%). PRV-1 was overexpressed in 25/58 ET (43.1%), 35/37 PV (94.6%) and 6/8 IMF (75%) and EECs formation was seen in 31/58 ET (53.4%), 34/37 PV (91.9%) and 6/8 IMF (75%). All markers were simultaneously positive (group A) in 43/103 patients (41.7%), concurrently negative (group B) in 25/103 patients (24.3%) and both positive and negative markers (group C) were observed in 35/103 patients (34%). In group A, 65% were PV patients, 26% were ET patients and 9 % were IMF patients. In group B, 8% were PV patients, 84% were ET patients and 8 % were IMF patients. In group C, 20% were PV patients, 74% were ET patients and 6 % were IMF patients. Regarding diagnosis, 76% of PV and 50% of IMF patients belonged to group A, whereas the majority of ET patients (45%) pertained to group C (table). When comparing ET and PV, a significant difference was observed (p<0.001) concerning group distribution. Conclusion. These results show that, although a good correlation has been observed for the simultaneous expression of these three biomarkers, differences in the pattern of positivity, specially in ET, indicate that not all Ph-negative patients share the same pathogenetic mechanisms and point to other coexisting genetic abnormalities. Diagnosis ET PV IMF All markers positive (group A) 11 (19%) 28 (76%) 4 (50%) All markers negative (group B) 21 (36%) 2 (5%) 2 (25%) Positive and negative markers 26 (45%) 7 (19%) 2 (25%) Total 58 37 8

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Association of antiphospholipid antibodies with clinical activity and renal pathological activity in patients with lupus nephritis

Lupus ◽

10.1177/09612033211006781 ◽

2021 ◽

pp. 096120332110067

Author(s):

Jianna Zhang ◽

Ji Zhang ◽

Qiongxiu Zhou ◽

Guoyuan Lu ◽

Xiaohan You

Keyword(s):

Lupus Nephritis ◽

Antiphospholipid Antibodies ◽

Activity Index ◽

Serum Levels ◽

Anticardiolipin Antibodies ◽

Clinical Activity ◽

Negative Group ◽

Lower Serum ◽

Positive Group ◽

Glycoprotein I

Objectives This study aimed to investigate the association of antiphospholipid antibodies (aPL) with clinical activity and renal pathological activity in patients with lupus nephritis (LN). Methods Levels of anticardiolipin () antibodies, anti-β2-glycoprotein I (anti-β2-GPI) antibodies and lupus anticoagulant (LAC) were measured, and other clinical and pathological data were also obtained during the same period before renal biopsy. Results A total of 83 patients with LN were included in this study, 40 patients (48.2%) in the s positive group and 43 patients in the aPL negative group. LN patients with positive aPL had significantly higher SLEDAI (p = 0.012), more hematuria (p = 0.043), lower serum C3 (p = 0.003) and C4 (p = 0.014), and a higher pathological activity index (p = 0.012), more micro-thrombosis (p = 0.046) and more C3 deposits (p = 0.038) in the glomerulus than patients with negative aPL The level of IgG- was significantly correlated with SLEDAI and serum level of C3 (r = 0.44, p < 0.001; r = −0.39, p = 0.003, respectively). The level of IgM- was significantly correlated with SLEDAI, and serum levels of C3 and C4 (r = 0.27, p = 0.014; r = −0.22, p = 0.041; r = −0.23, p = 0.035, respectively). Conclusions Our work suggests that aPL, especially, are correlated with both clinical activity and renal pathological activity in patients with LN.

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Characteristics of the Interaction between Thrombin Exosite1 and the Sequence 269-297 of Platelet Glycoprotein Ibα

Thrombosis and Haemostasis ◽

10.1055/s-0037-1615193 ◽

1998 ◽

Vol 80 (08) ◽

pp. 310-315 ◽

Cited By ~ 8

Author(s):

Marie-Christine Bouton ◽

Christophe Thurieau ◽

Marie-Claude Guillin ◽

Martine Jandrot-Perrus

Keyword(s):

Platelet Activation ◽

Electrostatic Interactions ◽

Platelet Glycoprotein ◽

Thrombin Receptor ◽

Central Position ◽

Amidolytic Activity ◽

N Terminus ◽

Hydrolysis Of ◽

Chemical Cross Linking ◽

Positively Charged

SummaryThe interaction between GPIb and thrombin promotes platelet activation elicited via the hydrolysis of the thrombin receptor and involves structures located on the segment 238-290 within the N-terminal domain of GPIbα and the positively charged exosite 1 on thrombin. We have investigated the ability of peptides derived from the 269-287 sequence of GPIbα to interact with thrombin. Three peptides were synthesized, including Ibα 269-287 and two scrambled peptides R1 and R2 which are comparable to Ibα 269-287 with regards to their content and distribution of anionic residues. However, R2 differs from both Ibα 269-287 and R1 by the shifting of one proline from a central position to the N-terminus. By chemical cross-linking, we observed the formation of a complex between 125I-Ibα 269-287 and α-thrombin that was inhibited by hirudin, the C-terminal peptide of hirudin, sodium pyrophosphate but not by heparin. The complex did not form when γ-thrombin was substituted for α-thrombin. Ibα 269-287 produced only slight changes in thrombin amidolytic activity and inhibited thrombin binding to fibrin. R1 and R2 also formed complexes with α-thrombin, modified slightly its catalytic activity and inhibited its binding to fibrin. Peptides Ibα 269-287 and R1 inhibited platelet aggregation and secretion induced by low thrombin concentrations whereas R2 was without effect. Our results indicate that Ibα 269-287 interacts with thrombin exosite 1 via mainly electrostatic interactions, which explains why the scrambled peptides also interact with exosite 1. Nevertheless, the lack of effect of R2 on thrombin-induced platelet activation suggests that proline 280 is important for thrombin interaction with GPIb.

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Platelet Activation and Aggregation Induced by Recombinant von Willebrand Factors Reproducing Four Type 2B von Willebrand Disease Missense Mutations

Thrombosis and Haemostasis ◽

10.1055/s-0037-1614241 ◽

1998 ◽

Vol 79 (01) ◽

pp. 211-216 ◽

Cited By ~ 13

Author(s):

Lysiane Hilbert ◽

Claudine Mazurier ◽

Christophe de Romeuf

Keyword(s):

Platelet Aggregation ◽

Platelet Activation ◽

Von Willebrand Disease ◽

Platelet Glycoprotein ◽

Missense Mutations ◽

Inhibit Platelet Aggregation ◽

Wild Type ◽

A1 Domain ◽

Von Willebrand ◽

Willebrand Factor

SummaryType 2B of von Willebrand disease (vWD) refers to qualitative variants with increased affinity of von Willebrand factor (vWF) for platelet glycoprotein Ib (GPIb). All the mutations responsible for type 2B vWD have been located in the A1 domain of vWF. In this study, various recombinant von Willebrand factors (rvWF) reproducing four type 2B vWD missense mutations were compared to wild-type rvWF (WT-rvWF) for their spontaneous binding to platelets and their capacity to induce platelet activation and aggregation. Our data show that the multimeric pattern of each mutated rvWF is similar to that of WT-rvWF but the extent of spontaneous binding and the capacity to induce platelet activation and aggregation are more important for the R543Q and V553M mutations than for the L697V and A698V mutations. Both the binding of mutated rvWFs to platelets and platelet aggregation induced by type 2B rvWFs are inhibited by monoclonal anti-GPIb and anti-vWF antibodies, inhibitors of vWF binding to platelets in the presence of ristocetin, as well as by aurin tricarboxylic acid. On the other hand, EDTA and a monoclonal antibody directed against GPIIb/IIIa only inhibit platelet aggregation. Furthermore, the incubation of type 2B rvWFs with platelets, under stirring conditions, results in the decrease in high molecular weight vWF multimers in solution, the extent of which appears correlated with that of plasma vWF from type 2B vWD patients harboring the corresponding missense mutation. This study supports that the binding of different mutated type 2B vWFs onto platelet GPIb induces various degrees of platelet activation and aggregation and thus suggests that the phenotypic heterogeneity of type 2B vWD may be related to the nature and/or location of the causative point mutation.

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Plasma Thrombospondin as an Indicator of Intravascular Platelet Activation in Patients with Vasculitis

Thrombosis and Haemostasis ◽

10.1055/s-0038-1646003 ◽

1987 ◽

Vol 58 (03) ◽

pp. 850-852 ◽

Cited By ~ 15

Author(s):

M B McCrohan ◽

S W Huang ◽

J W Sleasman ◽

P A Klein ◽

K J Kao

Keyword(s):

Platelet Activation ◽

Plasma Levels ◽

Half Life ◽

Degradation Products ◽

Plasma Concentrations ◽

Primary Source ◽

Positive Correlation ◽

Activation Marker ◽

Fibrin Degradation Products ◽

The Mean

SummaryThe use of plasma thrombospondin (TSP) concentration was investigated as an indicator of intravascular platelet activation. Patients (n = 20) with diseases that have known vasculitis were included in the study. The range and the mean of plasma TSP concentrations of patients with vasculitis were 117 ng/ml to 6500 ng/ml and 791±1412 ng/ml (mean ± SD); the range and the mean of plasma TSP concentrations of control individuals (n = 33) were 13 ng/ml to 137 ng/ml and 59±29 ng/ml. When plasma TSP concentrations were correlated with plasma concentrations of another platelet activation marker, β-thromboglobulin (P-TG), it was found that the TSP concentration inei eased exponentially as the plasma β-TG level rose. A positive correlation between plasma levels of plasma TSP and serum fibrin degradation products was also observed. The results suggest that platelets are the primary source of plasma TSP in patients with various vasculitis and that plasma TSP can be a better indicator than β-TG to assess intravascular platelet activation due to its longer circulation half life.

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Linkage between H. pylori Infection and TNF-α polymorphism in The Pregnant Women

International Journal of Research in Pharmaceutical Sciences ◽

10.26452/ijrps.v9ispl1.1298 ◽

2018 ◽

Vol 9 (SPL1) ◽

Author(s):

Ghaidaa Raheem Lateef ◽

Azhar Omaran Al-Thahab

Keyword(s):

Pregnant Women ◽

Serum Antibody ◽

Rapid Test ◽

Negative Group ◽

Positive Group ◽

Gynecology And Obstetrics ◽

Tnf Α ◽

Significant Difference ◽

Pcr Products ◽

H Pylori

A study was performed on 100 pregnant women in the outpatient department of gynecology and obstetrics of Maternity and Children Hospital in Al-Diwaniya City during the period between (March to September 2016). One hundred blood samples (50 for patients and 50 for control) were collected under the supervision of the treating gynecologist. The detection of Helicobacter. pylori was done by the use of the serum antibody Rapid test. The results showed that 50 (100%) were positive and 50 (100%) were negative for H. pylori in above method.All blood of patients and control samples were used for the extraction of genomic DNA,where the 107 bp PCR product size. Genotyping of the TNF-α-308 SNP (G/A)was performed by restriction fragment length polymorphism PCR (RFLP-PCR). PCR products were digested with restr NcoI iction enzyme. Individuals with the TNF-α-308(GG) homozygote produced digested DNA bands at 80,and 20 bp bp. A heterozygous genotype ofTNF-α-308 (GA)produced 107 bp,80 bp,and 20 bp bands. Individuals with the TNF-α-308 (AA) homozygote genotype had no amplicon digested and generated only one band of 107 bp. There was a significant difference in the frequency of the TNF-α-308(GG)genotype between H. pylori positive group and H. pylori negative group(72%,78% respectively). Also for GA genotype,there was a significant difference between H. pylori positive group and H. pylori negative group(24%,18% respectively). Concerning the frequency of the TNF-α-308 (AA)genotype between H. pylori positive group and H. pylori negative group,there was no significant difference between the two groups.

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