Iraqi J Pharm Sci, Vol.23(1) 2014 H. pylori IgG , IgA and CA19-9 and CEA in Gastritis 68 Detection of Helicobacter Pylori IgG and IgA , Serum Biomarkers CA19-9 and CEA in Patients with Gastrointestinal Diseases Shurooq,R. Kadhim* ,1 , Ishraq,H. Alaiwi* and Rasha,R.** * Department of Clinical Laboratory Sciences, College of Pharmacy, Al-Mustansiriyah University ,Baghdad. Iraq. ** Ministry of Health Abstract: Gastrointestinal diseases and especially chronic gastritis are mainly induced by Helicobacter pylori infection, and provides the basis for gastric carcinogenesis and colorectal cancer. The study involved the detection of serum anti-H. pylori IgG and IgA antibody of and some serum biomarkers ;CEA and CA19-9 in patients with gastrointestinal diseases. Fifty eight serum samples were collected from 25 males and 33 females .Peripheral venous blood was collected from each patient and sera obtained by centrifugation. Serum anti-H. pylori IgG and IgA ,serum CEA and CA19-9 were evaluated by enzyme-linked immunoadsorbent assays (ELISA).Forty eight serum samples were positive for IgG (82.7% ) divided into 27 positive samples for females and 21 positive samples for males while ten samples were positive for IgA ( 17.2%) divided into eight positive samples for females and two positive samples for males .Only three samples were positive for anti IgG and serum biomarker CA19-9 .In this study most of positive results of H.Pylori IgA and IgG are between the age 30- 50 years and low within 1-10 years ,we conclude because most of them are working and eat fast food from different sources. This study concluded that detection of serum IgG and IgA is effective in monitoring the H. pylori infection in gastrointestinal disorders and chronic infections . Keywords: Gastrointestinal diseases, Gastritis , H.Pylori, IgG ,IgA ,CEA,CA19-9. CA19-9و المعلماث البُىلىجُت Helicobacter Pyloriلبكترَا IgA و IgGالتحرٌ عن اضداد الجهاز الهضمٍلدي مرضً CEA و شروق رَس كاظم * ،1 رشا رحُم** واشراق حسن علُىٌ* ، . انعزاق ،ثغذاد ،انجبيعخ انًسزُصزٌخ ،كهٍخ انصٍذنخ ، السريريةختبرية مفرع العلوم ال* . انعزاق ،ثغذاد ،** ٔسارح انصحخ الخالصة انًعذح ٔانًؤدي انى األصبثخ ثسزطبٌانشيٍ انًعذح يزاض انجٓبس انٓضًً ٔانزٓبةانًسجت انزئٍسً أل Helicobacter pyloriرعذ ثكززٌب ٔ CA19-9ٔانًعهًبد انجٍٕنٕجٍخ H. pylori ثكززٌب IgG ٔIgA انكهٕثٍٕنٍُبد انًُبعٍخ انزحزي عٍرضًُذ ْذِ انذراسخ .ٔانقٕنٌٕ CEA عٍُخ يٍ انذكٕر 58ٔرٕسعذ انى انشيٍانًعذح انجٓبس انٓضًً ٔانزٓبة عٍُخ يصم يٍ يزضى 85جًعذ . فً يصٕل انًزضى . فً يصٕل انًزضى ثطزٌقخ األنٍشا CA19-9 ٔCEAٔانًعهًبد انجٍٕنٕجٍخ IgG ٔ IgA عٍُخ يٍ األَبس ٔقٍسذ اضذاد ال 33ٔ عٍُخ نألَبس . ثًٍُب كبَذ انعٍُبد انًٕججخ ألضذاد 52عٍُخ نهذكٕر ٔ 52%.( رٕسعذ ال 55.5)IgG عٍُخ اٌجبثٍزٓب ألضذاد ال 85اظٓزد IgA (22.5ٕرٕسعذ ثٍٍ ثًبٍَخ عٍُبد نألَبس ٔ اثُبٌ نهذك )%. ألضذاد ال أٔضحذ انُزبئج ظٕٓر ثالس عٍُبد يٕججخ ر IgG ٔانًعهى سُخ ألَّ ْذِ انفئخ ًْ األكثز َشبطب ٔاررجبطب 83-33نألعًبر IgG ٔ IgA.كبَذ َزبئج ْذِ انذراسخ اٌجبثٍخ ألضذاد CA19-9انجٍٕنٕجً سُٕاد ًْ األقم 23-2ثبنعًم ٔاألكثز رعزضب نٓذِ انجكززٌب ألٌ يعظًٓى ٌزُبٔنٌٕ انٕججبد انسزٌعخ ٔيٍ يصبدر يخزهفخ.ثًٍُب كبَذ انفئخ انعًزٌخ ثبسزخذاو انفحص انسٍزٔنٕجً ٔ انذي ْٕ يٍ انفحٕصبد انزٔرٍٍُخ H. pylori جكززٌبن IgG ٔ IgAاصبثخ . ٌعذ انزحزي عٍ األجسبو انًضبدح ٔانًزٕفزح فعبال فً انزحزي عٍ األصبثخ ثٓذِ انجكززٌب خصٕصب فً األصبثبد انًشيُخ ٔاألضطزاثبد انًعٌٕخ. . السمنالمعدة أمراض الجهاز الهضمٍ والتهاب ، H. pylori، IgG ، IgA ،CA19-9 ، CEA الكلماث المفتاحُت: Introduction Helicobacter pylori , a spiral - shaped pathogenic bacterium found on the human gastric mucosa , was first isolated by Warren and Marshall (1) in 1982 and soon after was Linked with chronic gastritis and peptic ulceration (2) . The world wide prevalence of H. pylori is more than 50% (3,4) . It is more prevalent in developing countries as compared to developed countries (5) . Its prevalence in South Asia is ranging between 55 to 90% (6) . Primary care physicians were reported that 78% of the physicians thought that contaminated water was the source of spread of infection (7) Chronic gastritis is mainly induced by Helicobacter pylori infection, and provides the basis for gastric carcinogenesis (8) . Several prospective studies reported a strong association between H. pylori infection and gastric cancer; hence , H. pylori is now recognized as a Group 1 carcinogen for humans (9,10) . However , only a few individuals 1 Corresponding author E-mail: shrooq7@yahoo.com Received: 20/3/2013 Accepted: 22/4/2014 Iraqi J Pharm Sci, Vol.23(1) 2014 H. pylori IgG , IgA and CA19-9 and CEA in Gastritis 69 infected by H. pylori may eventually develop gastric cancer. The clinical outcome of H. pylori infection may depend on several factors, such as age at the time of first infection, environmental factors, immune response, and genetic characteristics (11) .Among the serum tumor markers, CEA and CA19-9 are most widely used for the screening and monitoring gastric cancer, because they have been reported to be elevated in some patients with gastric cancer (12,13) . Carcinoembryonic antigen (CEA) is the most commonly used tumor marker in patients with colorectal cancer, and the tumor marker carbohydrate antigen CA19- 9 is often used in combination with CEA to manage patients with colorectal cancer (14,15,16) . The aim of the study is to determine the prevalence of the serum anti-H. pylori immunoglobulin IgG and IgA antibodies and serum tumor markers, CEA and CA19-9 in patients with gastrointestinal diseases . Patients and Methodes Patients and samples: A total of fifty eight patients with abdominal pains, cramps, diarrhea and chronic gastritis whom referred to Al-Karamah hospital in Baghdad from April 2010 to July 2011, were included in the present study . Among them 25 were males and 33 were females with gastrointestinal disorders ( 8 persons were considered as control) , aged from 6 to 72 years with a mean age of 55( - + )years old. Serological tests: Blood was sampled twice from patients. Enzyme-linked immunoadsorbent assays (ELISA) were used to measure the levels of serum anti-H. pylori IgG and IgA antibodies. The test kits for the detection of anti-H. pylori -IgG and anti- H.pylori IgA were purchased by Biohit Oyi company (Finland) . Detection of serum markers CEA and CA19-9 were also done by ELISA technique (Biohit Oyi company) . Results and Discussion The results were classed as positive if anti–H pylori immunoglobulin IgG titers were >30 U/mL and IgA titers positive if they were >20 U/mL. Table 1 shows the detective positive rates of serum anti H. pylori IgG and IgA antibodies of patients in relation to gender and age. The incidence of infection was (43.18 %, 25/58) between the males and (56.8 %, 33/58) the females patients .The H. pylori infection rate was the highest in the ages from 42 to 50 years.Also the detected positive rate of H. pylori was not remarkably associated with the age of patients . Table (1) Age ,gender and number of anti H.pylori IgA and IgG positive serum samples in females and males. The results showed that anti H.pylori IgG positive serum samples in both females and males were elevated in ages 21-50 years as in table 2 and 3. Table (2) Age distribution of anti H.pylori IgG positive serum samples in females Age (year) Gender Numbers of patients 1-10 F 2 11-20 = 3 21-30 = 5 31-40 = 8 41-50 = 7 51-60 = 2 Total = 27 Table (3)Age distribution of anti H.pylori IgG positive serum samples in males Anti H.pylori IgA positive serum samples in males were only 2 between ages 1-30 years as in table 4,but anti H.pylori IgA positive serum samples in females were elevated in ages 41- 50 years as in table 5. Gender Age (year ) Number and percentage of patients with positive anti -IgA Numbers and percentage of patients with positive anti -IgG F 7-60 8(13.7%) 27(46.5%) M 1-58 2(3.4%) 21(36.2%) 10(17.1%) 48(82.7%) Age (year) Gender Numbers of patients 1-10 M 1 11-20 = 1 21-30 = 4 31-40 = 5 41-50 = 7 51-60 = 3 Total = 21 Iraqi J Pharm Sci, Vol.23(1) 2014 H. pylori IgG , IgA and CA19-9 and CEA in Gastritis 70 Table (4 ) Age distribution of anti H.pylori IgA positive serum samples in males Age (year) Gender Numbers of patients 1-10 M 1 11-20 = - 21-30 = 1 31-40 = - 41-50 = - 51-60 = - Total 2 Table ( 5 ) Age distribution of anti H.pylori IgA positive serum samples in females Age (year) Gender Numbers of patients 1-10 F - 11-20 = - 21-30 = 1 31-40 = 1 41-50 = 6 51-60 = - Total 8 Table (6 ) Age distribution of anti H.pylori IgG and IgA positive serum samples in females . Age (year) Gender Numbers of positive IgG and IgA samples 1-10 F - 11-20 = - 21-30 = - 31-40 = 1 41-50 = 3 51-60 = 1 Total 5 Table (7 )Age distribution of anti H.pylori IgG and IgA positive serum samples in males . Table (8 ) Age distribution of anti H.pylori IgG and IgA positive and serum markers CEA and CA19-9 in males patients . Three specimens of males gave positive result for CA19-9 test but all females specimens gave negative results for CA19-9 test. All females and males specimens gave negative results for CEA test. It is now recognized that the clinical detection of serum antibody is effective in monitoring the H. pylori infection and the serology test has been used widely in epidemiologic studies and routine diagnosis of H pylori infection (5,6 ,7,8) .In this study most of positive results of IgA and IgG are between the age 30-50 years and low within 1-10 years ,we conclude because most of them are working and eat fast food from different sources. Serological testing to diagnose H. pylori infection in children is still controversial, although commonly used in clinical practice. A recent study speculate that young children may have a different immune response to H. pylori, with preferable responses to certain antigens, as well as lower titers than adults. The Pyloriset test may fail to recognize these specific antibodies (17) .However, most individuals infected with H. pylori do not experience symptoms or have signs of recognizable disease. In most children, the presence of H. pylori infection does not lead to clinically apparent disease, even when the organism colonizing the gastric mucosa causes chronic active gastritis (18) . A recent study was carried in Iran and this study showed that IgG titers were high in individuals between 40 to 50 years old , Anti H. pylori IgA was also correlated with increasing age (19) . Serological tests are widely available and cheap, and may be helpful-in screening populations or in confirming the presence H. pylori infection in case of equivocal results of the other diagnostic methods due to bleeding ulcers, antibiotic and/or antisecretory treatments. A major advantage of this serologic test is that it Age (year) Gender Numbers of positive IgG and IgA samples 1-10 M 1 11-20 = - 21-30 = - 31-40 = 1 41-50 = - 51-60 = - Total 2 Iraqi J Pharm Sci, Vol.23(1) 2014 H. pylori IgG , IgA and CA19-9 and CEA in Gastritis 71 enables large numbers of subjects to be screened quickly and relatively inexpensively (20) .In table (8) the results showed only three males infected with H.pylori and elevated values of CA19-9 in age between 11-50 years. According to these results our study conclude that these three males may suffered from gastric or other kind of gastrointestinal tumors . A low number of positive results were recorded on elevated values of CA19-9 by Kim et al. when they identified only four patients among 1,063 patients with CA 19-9 serum levels >37 U/ml (mean values 50.5±16.8 U/ml) (21) .Some times CA19-9 is not used for early screening because it is not present in patients with certain blood types and is often elevated in benign disease. Certain changes that occur in the sera of pancreatic cancer patients reflect the high level of inflammation associated with the disease (22) . A recent study on serum CA19-9 mention that it is the most extensively studied and clinically useful biomarker for pancreatic cancer. Unfortunately, CA 19-9 serum level evaluation in pancreatic cancer patients is limited by poor sensitivity, false negative and increased false positivity in the presence of obstructive jaundice (10–60%) (23) . We conclude that serum analysis should include CA19-9 and CEA for the diagnosis the chronic infections with H. pylori. 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