scholarly journals Usefulness of Polymerase Chain Reaction in the diagnosis of asymptomatic malaria among school age children in Ilorin, Nigeria

2018 ◽  
Vol 1 (1) ◽  
pp. 39-45
Author(s):  
Ayodele Adedoja ◽  
Shola Kola Babatunde ◽  
Bukola Deborah Tijani ◽  
Ajibola A. Akanbi II ◽  
Olusola Ojurongbe
Keyword(s):  
Polymerase Chain Reaction ◽  
Plasmodium Species ◽  
Malaria Diagnosis ◽  
School Age Children ◽  
Clinical Settings ◽  
Asymptomatic Malaria ◽  
Chain Reaction ◽  
Malaria Species ◽  
Pcr Method ◽  
Polymerase Chain

Introduction: In Nigeria, microscopy and Rapid Diagnostic Tests (RDTs) are majorly used routinely to diagnose malaria in clinical settings. In recent years, molecular diagnosis has emerged as the most sensitive method for malaria diagnosis. This study determined the usefulness of Polymerase Chain Reaction (PCR) in comparison to microscopy (Giemsa stained thick and thin smears) for the diagnosis of asymptomatic malaria in Ilorin, Nigeria. Methods: Th e study enrolled 310 school children aged 4–15 years with no sign or symptoms of malaria. Blood samples were collected for identification of Plasmodium species infection using light microscopy and conventional PCR. Results: The PCR method detected m o r e infection of P. falciparum 107 (34.5%) than slide microscopy 81 (26.1%) in the study. Only P. falciparum was detected by microscopy while PCR detected mono infection of P. malariae (3.2%) and P. ovale (0.6%) and mixed infection of P. falciparum and P. malariae (3.2%). Overall the PCR method detected more malaria parasite compared to microscopy. Conclusion: The PCR technique, although more laborious and expensive than microscopy, have better diagnostic accuracy and are highly useful for the detection of P. falciparum and other malaria species in asymptomatic and low parasitaemia cases.

UJI DIAGNOSTIK POLYMERASE CHAIN REACTION DIBANDINGKAN DENGAN PENGECATAN GIEMSA PADA INFEKSI MALARIA

2018 ◽  
Vol 6 (1) ◽  
pp. 76-86
Author(s):  
Hanina Hanina
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Test ◽  
Sensitivity And Specificity ◽  
Predictive Value ◽  
Plasmodium Species ◽  
Malaria Diagnosis ◽  
Chain Reaction ◽  
Consecutive Sampling ◽  
Pcr Method ◽  
Polymerase Chain

ABSTRACT Plasmodium is a parasite causing malaria, the most important infection disease in the world. Gold standard of malaria diagnosis was founded Plasmodium by Giemsa staining method. Fundamental difference between Giemsa and Polymerase Chain Reaction (PCR) is the ability to detect parasite. Giemsa can detect minimal 50-100 parasit/μl whereas PCR detect parasite DNA in lower parasitemia. The purpose of this study was to determine the sensitivity and specificity of the PCR compared to blood slide with Giemsa in detecting of malaria infection. A diagnostic test has been conducted in Laboratorium Biomedik Fakultas Kedokteran Universitas Jambi and Laboratorium Biomolekuler Fakultas Kedokteran Universitas Sriwijaya. There were 87 subjects who fulfilled the criteria inclusion and drawn by consecutive sampling. Blood samples were taken from venous blood. Detection of Plasmodium used Giemsa and PCR method. Detection of Plasmodium from 87 subjects, Giemsa and PCR method founded 1 subject (1.1%) P. falciparum and 4 subjects (4.6%) P. vivax. 82 subjects (94.3%) were negative. The sensitivity and specificity of PCR was 100%, positive predictive value and negative predictive value  was 100%.Conclusion is higher sensitivity and spesificity PCR methode in the malaria diagnosis was proven and PCR methode able to identified Plasmodium species accuratly.   Keywords: Plasmodium, Malaria, Giemsa, PCR, Diagnostic Test   ABSTRAK Plasmodium merupakan parasit penyebab malaria, suatu penyakit infeksi paling penting di dunia. Baku emas diagnosa malaria  adalah menemukan Plasmodium melalui pemeriksaan mikroskopis dengan pengecatan Giemsa. Perbedaan mendasar antara metode Giemsa dengan metode Polymerase Chain Reaction (PCR) terletak pada kemampuan mendeteksi parasit. Metode Giemsa hanya mampu mendeteksi Plasmodium dengan ambang batas antara 50-100 parasit/μl sedangkan metode  PCR dapat mendeteksi DNA parasit pada parasitemia yang lebih rendah. Tujuan penelitian ini adalah untuk mengetahui sensitivitas dan spesifisitas metode PCR dibandingkan dengan pengecatan Giemsa dalam menegakkan diagnosis infeksi malaria. Penelitian ini merupakan uji diagnostik yang dilakukan di Laboratorium Biomedik Fakultas Kedokteran dan Ilmu Kesehatan Universitas Jambi dan Laboratorium Biomolekuler Fakultas Kedokteran Universitas Sriwijaya. Penelitian dilakukan mulai bulan Januari sampai April 2017. Subjek penelitian berjumlah 87 orang yang diambil secara consecutive sampling dari laboratorium RS Theresia Jambi. Semua subjek diambil sampel darah venanya, kemudian dilakukan pengecatan Giemsa dan pemeriksaan PCR. Hasil pemeriksaan PCR nested menggunakan primer genus dan primer spesies Plasmodium ditemukan P.falciparum positif sebanyak 1 sampel (1,1%). Sedangkan P.vivax positif sebanyak 4 sampel (4,6%). Sebanyak 82 sampel (94,3%) negatif. Hal ini sesuai dengan hasil pemeriksaan mikroskopis dengan pengecatan Giemsa. Metode PCR dibandingkan dengan metode pengecatan Giemsa sensitivitas dan spesifisitasnya 100%, nilai prediksi positif dan nilai prediksi negatifnya 100%. Dapat disimpulkan bahwa metode PCR sangat sensitif dan spesifik dalam penegakan diagnosis malaria dan mampu mengidentifikasi spesies parasit secara akurat.   Kata Kunci: Plasmodium, Malaria, Giemsa, PCR, Uji Diagnostik.

scholarly journals How prevalent is Plasmodium malariae in Rondônia, Western Brazilian Amazon?

2000 ◽  
Vol 33 (5) ◽  
pp. 489-492 ◽  
Author(s):  
Marisa Torres Vidal Cavasini ◽  
Weber Luidi Ribeiro ◽  
Fumihiko Kawamoto ◽  
Marcelo Urbano Ferreira
Keyword(s):  
Polymerase Chain Reaction ◽  
Plasmodium Species ◽  
Brazilian Amazon ◽  
Plasmodium Malariae ◽  
Mixed Species ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Potential Impact ◽  
And Control

We have compared results of Plasmodium species identification obtained with conventional on-site microscopy of Giemsa-stained thick smears (GTS) and a semi-nested polymerase chain reaction (PCR) in 96 malaria patients from Rondônia, Western Brazilian Amazon. Mixed-species infections were detected by PCR in 30% patients, but no such case had been found on GTS. Moreover, P. malariae infections were detected in 9 of 96 patients (10%) by PCR, but were not identified by local microscopists. The potential impact of species misidentification on malaria treatment and control is discussed.

scholarly journals PCR - based diagnosis to evaluate the performance of malaria reference centers

2004 ◽  
Vol 46 (4) ◽  
pp. 183-187 ◽  
Author(s):  
Silvia Maria Di Santi ◽  
Karin Kirchgatter ◽  
Karen Cristina Sant'Anna Brunialti ◽  
Alessandra Mota Oliveira ◽  
Sergio Roberto Santos Ferreira ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Gold Standard ◽  
Blood Smear ◽  
Plasmodium Species ◽  
Thick Blood Smear ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Positive Results ◽  
Ssrrna Gene

Although the Giemsa-stained thick blood smear (GTS) remains the gold standard for the diagnosis of malaria, molecular methods are more sensitive and specific to detect parasites and can be used at reference centers to evaluate the performance of microscopy. The description of the Plasmodium falciparum, P. vivax, P. malariae and P. ovale ssrRNA gene sequences allowed the development of a polymerase chain reaction (PCR) that had been used to differentiate the four species. The objective of this study was to determine Plasmodium species through PCR in 190 positive smears from patients in order to verify the quality of diagnosis at SUCEN's Malaria Laboratory. Considering only the 131 positive results in both techniques, GTS detected 4.6% of mixed and 3.1% of P. malariae infections whereas PCR identified 19.1% and 13.8%, respectively.

scholarly journals Diversity ofLeptospiraspp. in Rats and Environment from Urban Areas of Sarawak, Malaysia

2017 ◽  
Vol 2017 ◽  
pp. 1-8 ◽  
Author(s):  
Chai Fung Pui ◽  
Lesley Maurice Bilung ◽  
Kasing Apun ◽  
Lela Su’ut
Keyword(s):  
Polymerase Chain Reaction ◽  
Water Samples ◽  
Urban Areas ◽  
Environmental Samples ◽  
Soil Samples ◽  
Chain Reaction ◽  
Prevalence Studies ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Soil And Water

Various prevalence studies onLeptospirain animals and humans, as well as environmental samples, had been conducted worldwide, including Malaysia. However, limited studies have been documented on the presence of pathogenic, intermediate, and saprophyticLeptospirain selected animals and environments. This study was therefore conducted to detectLeptospiraspp. in rats, soil, and water from urban areas of Sarawak using the polymerase chain reaction (PCR) method. A total of 107 rats, 292 soil samples, and 324 water samples were collected from April 2014 to February 2015. PathogenicLeptospirawas present in 5.6% (6/107) of rats, 11.6% (34/292) of soil samples, and 1.9% (6/324) of water samples. IntermediateLeptospirawas present in 2.7% (8/292) of soil samples and 1.9% (6/324) of water samples. SaprophyticLeptospirawas present in 10.3% (11/107) of rats, 1.4% (4/292) of soil samples, and 0.3% (1/324) of water samples. From this study, 76Leptospiraspp. were isolated. Based on DNA sequencing, the dominantLeptospiraspp. circulating in urban areas of Sarawak are pathogenicLeptospira noguchii, intermediateLeptospira wolffiiserovar Khorat, and saprophyticLeptospira meyeri, respectively. Overall, this study provided important surveillance data on the prevalence ofLeptospiraspp. from rats and the environment, with dominant local serovars in urban areas of Sarawak.

Quantitative Real-Time Polymerase Chain Reaction Detection of BK Virus Using Labeled Primers

2010 ◽  
Vol 134 (3) ◽  
pp. 444-448 ◽  
Author(s):  
Zhengming Gu ◽  
Jianmin Pan ◽  
Matthew J. Bankowski ◽  
Randall T. Hayden
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Real Time Pcr ◽  
Quantitative Polymerase Chain Reaction ◽  
Bk Virus ◽  
Clinical Samples ◽  
Quantitative Detection ◽  
Chain Reaction ◽  
Pcr Method ◽  
Polymerase Chain

Abstract Context.—BK virus infections among immunocompromised patients are associated with disease of the kidney or urinary bladder. High viral loads, determined by quantitative polymerase chain reaction (PCR), have been correlated with clinical disease. Objective.—To develop and evaluate a novel method for real-time PCR detection and quantification of BK virus using labeled primers. Design.—Patient specimens (n = 54) included 17 plasma, 12 whole blood, and 25 urine samples. DNA was extracted using the MagNA Pure LC Total Nucleic Acid Isolation Kit (Roche Applied Science, Indianapolis, Indiana); sample eluate was PCR-amplified using the labeled primer PCR method. Results were compared with those of a user-developed quantitative real-time PCR method (fluorescence resonance energy transfer probe hybridization). Results.—Labeled primer PCR detected less than 10 copies per reaction and showed quantitative linearity from 101 to 107 copies per reaction. Analytical specificity of labeled primer PCR was 100%. With clinical samples, labeled primer PCR demonstrated a trend toward improved sensitivity compared with the reference method. Quantitative assay comparison showed an R2 value of 0.96 between the 2 assays. Conclusions.—Real-time PCR using labeled primers is highly sensitive and specific for the quantitative detection of BK virus from a variety of clinical specimens. These data demonstrate the applicability of labeled primer PCR for quantitative viral detection and offer a simplified method that removes the need for separate oligonucleotide probes.

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