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Development of an Indigenous Real Time Polymerase Chain Reaction for the Diagnosis of Malaria

Thesis Info

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Author

Khan, Dr. Saleem Ahmed

Program

PhD

Institute

Riphah International University

City

Islamabad

Province

Islamabad

Country

Pakistan

Thesis Completing Year

2013

Thesis Completion Status

Completed

Subject

Chemistry

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/2480/1/3080S.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725867690

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INTRODUCTION Establishing highly sensitive, affordable diagnostic test covering all species of malaria is needed in countries like Pakistan that has significant prevalence of this disease. Conventional tests, like microscopy are widely used but it has certain performance related limitations. Rapid diagnostic tests (RDTs) for plasmodium have not achieved greater sensitivity than microscopy. Polymerase chain reaction (PCR) based methods have shown lot of promise and these are replacing microscopy as gold standard. OBJECTIVES To establish a real time PCR for malaria diagnosis and to compare its accuracy with microscopy and an antigen based RDT. STUDY DESIGN AND SETTING This cross sectional analytical study was conducted at Military Hospital and Armed forces Institute of Pathology Rawalpindi, Pakistan from January 2011 to December 2011. SUBJECTS The study included 300 clinically suspected patients of malaria. 3 METHODS Venous blood sample from each patient was tested for malarial parasite by microscopy and antigen detection assay (OptiMal). Parasite index was calculated. Plasmodium genus specific in house real time PCR, on all specimens was performed targeting small subunit rRNA gene. Diagnostic accuracy of three tests were compared and cost analysis done. RESULTS Out of 300 patients malaria parasite was detected in 110, 106 and 123 patients by microscopy, OptiMAL and PCR respectively. Real time PCR was 100% sensitive while microscopy and OptiMal had sensitivity of 89.4% and 86.2% respectively. All methods were 100% specific. The cost per test was 0.2, 2.75 and 3.30 US$ by microscopy, OptiMal and PCR respectively, excluding the once capital cost on PCR equipment. Conclusion: Real time PCR for diagnosis of malaria was successfully established. The technique is highly sensitive and affordable. It is recommended to be incorporated in the diagnostic algorithm for malaria in Pakistan. Key Words: Malaria, Microscopy, Rapid Diagnostic Tests (RDTs), Polymerase Chain Reaction (PCR).
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تقریظ سوئم

تقریظ سوئم
بسم اللہ الرحمن الرحیم
نحمدہ ونصلی علی رسولہِ الکریم

مُوصوف ایک علمی گھرانے سے تعلق رکھتے ہیں۔ دینی علوم کے حوالے سے ان کے بزرگوں کی خدمات اظہر من الشمس ہیں ۔اِن کی کتاب ــ ’’ نگارشاتِ راشد‘‘ ایک علمی خزانہ ہے اِس میں طلباء و طالبات کے لیے متعدد مضامین شامل کی گئی ہیں۔ جو اپنی نظیرآپ ہیں حافظ محمد اکرم راشدؔ نے دیگر کتب اور مضامین سے خوشہ چینی کی بجائے فی البدیہہ مواد پیش کرنے کی مساعی جمیلہ کی ہے ۔ اِ ن کی تحریر میں چاشنی اور ندرت ہے۔ اِن کے مضامین جو’’ نگارشاتِ راشد ‘‘کے نام سے زیورِ طباعت سے مرصعّ ومزیّن ہو رہے ہیں عام قاری کے لیے بالعموم اور طلباء و طالبات کے لیے بالخصوص ممدو معاون ثابت ہوں گے۔ ادارہ کے مقابلہ جات ہوں یا ضلعی اور ڈویژن لیول کے مقابلہ جات ’’ نگارشاتِ راشد‘‘ میں شامل شدہ مضامین کفایت کریں گے ۔ یہ طلباء کے لیے نعمت مترقبہ سے کم نہ ہے ۔ موصوف کا ادبی دنیا میں ایک نام ہے اِ ن کو دیکھ کر متقدمین ادباء کی یاد تازہ ہو جاتی ہے ۔ دورانِ ملازمت بھی ادبی پروگرام کے حوالے سے محکمہ تعلیم میں اِن کی تحریریں محو گردش رہی ہیں ۔ اور انعام و اکرام کی حقدار گردانی گئی ہیں۔
اللہ تعالیٰ اِن کو مزید ایسے شاہ پارے پیش کرنے کی توفیق عطا فرمائے ۔
رانا محمد اظہر خاں
چیف ایگزیکٹو آفیسر (ایجوکیشن)
ڈسٹرکٹ ایجوکیشن اتھار ٹی ، رحیم یار خاں

Challenges of Translating the Arabic Qur’an into English A Comparative Study of Eight Leading Translations

Any translation of the Arabic Qur’an in English or any European language is likely to be imperfect. This is primarily due to the differences in the language, semantics, idiom, style and culture. Almost fifty such translations have appeared in the last fifty years, both by Muslim and other scholars, but none can claim any perfection in imaging the Arabic Qur’an. Nevertheless, there are some that are faithful to word-by-word (literal) or sense-for-sense (free) translation, but most lack the flavor of the Qur’anic essence and image either due to the translating approach, or inadequate understanding of the meaning of Sacred Arabic Text, or constraints of eloquence of the English language. This paper examines eight of the leading translations and draws conclusions relating to the use of translation techniques and literary devices and concepts that add beauty to the eloquence of Arabic Qur’an and makes it a living and literary masterpiece. It is found that the meaning of the lexical expressions have been maintained to a high degree in the process of translation and the use of literary devices has been adequately captured by the selected translations.

Role of Halocarbons and Polychlorinated Biphenyls in the Development of Hepatitis

The present work is concerned with the determination of blood parameters, enzyme levels, blood chemistry and pesticide residues in the blood samples from different areas of Karachi in comparison with unexposed persons (i.e., control). Total 45 persons (exposed to pesticides) were taken in this study to assay the pesticides residues in their blood and also observed the morphological and chemical changes in the blood of affected persons. From 45 persons, only 20 persons were found positive and included in the study. Out of ten (10) standard pesticides five differentpesticides were identified in the blood of exposed persons such as cypermethrin, deltamethrin, polytrin-C, malathion and permethrin. The other standard pesticides such as DDE, DDT, match, diazinon and monocrotophos were not detected. In addition blood samples were also collected from ten (10) normal persons (not exposed to pesticides) for the determination of blood parameters, enzymes levels and blood chemistry. Regarding blood parameters approximately all the blood samples had significant variations i.e., (increase or decrease level) possibly due to the cumulative effect of pesticide residues e.g., Majid a young male aged 25 years had been exposed to pesticides for long time and therefore his TLC and platelet counts were found high in addition to low levels of RBC count, Hb, Hct and MCH which probably due to the excessive secretion of enzymes. Similarly, Ghulam Hussain another young male person aged only 08 years had low levels of RBC, Hb, Hct, platelet, MCV and MCH, which may be due to toxic effects of pesticides. In differential leukocyte count (DLC), granulocyte count was found in lower limits in 40 samples and showed normal limits in 05 samples. Lymphocyte count was xi found high in 31 samples. Low limit of lymphocyte was not found in any person. Similarly, monocyte count was found in high limits in 38 samples. Low monocyte count was not noted in any sample and the rest of the samples had normal limits of monocyte. Positive effects of pesticide residues were also observed on the enzyme levels. Regarding enzyme levels GPT, ALP, and ɣ - GT had higher levels in maximum number of samples. Cholinesterase was found high in 6 samples out of 45 samples and the highest was found in case of Farhan and Abdul Rahmanwhere as low limit of cholinesterase was noted in 15 samples out of total samples which may due to prolonged exposure of pesticides. GPT was found in high limits in all the samples. Also ɣ - GT and ALP found high in all the samples. Effect of pesticide residues were also observed on the blood chemistry. Regarding total bilirubin only one sample showed normal limit. The rest of the samples had raised total bilirubin and the highest was found in Javed a young male person aged 12 years. Similarly, in case of bilirubin direct all the blood samples showed increased level. The standard chromatograms of 10 different pesticides such as match, cypermethrin, deltamethrin, polytrin-C, diazinon, DDE, monocrotophos, DDT, malathion and permethrin were prepared on HPLC and their retention time (RT) was noted. Calculation was done for the quantity of pesticides residues from the peaks of chromatograms. Regarding pesticide residues, out of total 45 samples, only 20 persons were found positive for residues. The quantity of pesticide was determined by comparing xii the peak area of the standard pesticide with the sample peaks. After analysis, it was found that out of 20 samples deltamethrin and cypermethrin were found in noticeable amount in 6 samples. Also polytrin-C, malathion and permethrin were found in 2, 1 and 5 samples respectively. The pesticides such as match, diazinon, DDT and monocrotophoswere not detected in any sample.