scholarly journals Diagnostic test of urine sample, vaginal smear and combination of urine with vaginal smear to identify Neisseria gonorrhoeae with polymerase chain reaction method

2019 ◽  
Vol 7 (7) ◽  
pp. 2547
Author(s):  
Kristina Nadeak
Keyword(s):  
Polymerase Chain Reaction ◽  
Neisseria Gonorrhoeae ◽  
Diagnostic Test ◽  
Predictive Value ◽  
Reference Sample ◽  
Nucleic Acid Amplification ◽  
Urine Samples ◽  
Vaginal Smear ◽  
Chain Reaction ◽  
Polymerase Chain

Background: Gonorrhoea is a disease caused by Neisseria gonorrhoeae that is transmitted through sexual contact. There are several examinations performed on gonorrhoea infection, one of them is Polymerase Chain Reaction (PCR). The objective is to determine the diagnostic test of urine samples, vaginal smear and combination of urine and vaginal smear in identifying Neisseria gonorrhoeae using the PCR method.Methods: This study is a diagnostic test with a cross-sectional design involving 58 female sex workers (FSW). All FSWs are carried out of history and physical examination. Urine sampling, vaginal smear, combination of urine and vaginal smear, and endocervical smear were obtained for identifying Neisseria gonorrhoeae using PCR method, then a diagnostic test analysis of each sample was performed.Results: The diagnostic test of PCR for Neisseria gonorrhoeae from urine samples was found sensitivity 44.4%, specificity 20.0%, positive predictive value (PPV) 83.3%, negative predictive value (NPV) 3.8% and accuracy 42.0%. From vaginal smear, we obtained sensitivity 34.0%, specificity 66.7%, PPV 88.2%, NPV 12.1% and accuracy 38.0%. And from combination of urine and vaginal smear, we obtained sensitivity 51.1%, specificity 20.0%, PPV 85.2%, NPV 4.3% and accuracy 48.0%.Conclusions: From these results the researchers suggested that urine, vaginal and combination of urine and vaginal smear could not be used as an alternative to examine the sensitivity and specificity of Neisseria gonorrhoeae, so the endocervical sample remained the reference sample for examination of nucleic acid amplification tests for Neisseria gonorrhoeae.

scholarly journals Performance comparison of two malaria rapid diagnostic test with real time polymerase chain reaction and gold standard of microscopy detection method

2020 ◽  
Author(s):  
Puspa Wardhani ◽  
Trieva Verawaty Butarbutar ◽  
Christophorus Oetama Adiatmaja ◽  
Amarensi Milka Betaubun ◽  
Nur Hamidah ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Diagnostic Test ◽  
Real Time Pcr ◽  
Predictive Value ◽  
Gold Standard ◽  
Detection Method ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

Background: The diagnostic test for malaria is mostly based on Rapid Diagnostic Test (RDT) and detection by microscopy. Polymerase Chain Reaction (PCR) is also a sensitive detection method that can be considered as a diagnostic tool. The outcome of malaria microscopy detection depends on the examiner's ability and experience. Some RDT has been distributed in Indonesia, which needs to be evaluated for their results. Objective: This study aimed to compare the performance of RightSign RDT and ScreenPlus RDT for detection of Plasmodium in human blood. We used specific real-time polymerase chain reaction abTESTMMalaria qPCRII) and gold standard of microscopy detection method to measure diagnostic efficiency. Methods: Blood specimens were evaluated using RightSign RDT, ScreenPlus RDT, Microscopy detection, and RT-PCR as the protocol described. The differences on specificity (Sp), sensitivity (Sn), positive predictive value (PPV), and negative predictive value (NPV) were analyzed using McNemar and Kruskal Wallis analysis. Results: A total of 105 subjects were recruited. Based on microscopy test, RightSign RDT had sensitivity, Specificity, PPV, NPV, 100%, 98%, 98.2%, 100%, respectively. ScreenPlus showed 100% sensitivity, 98% specificity, 98.2% PPV, 100% NPV. The sensitivity of both RDTs became lower (75%) and the specificity higher (100 %) when using real-time PCR. Both RDTs showed a 100% agreement. RT-PCR detected higher mix infection when compared to microscopy and RDTs. Conclusion: RightSign and ScreenPlus RDT have excellent performance when using microscopy detection as a gold standard. Real-time PCR method can be considered as a confirmation tool for malaria diagnosis.

scholarly journals Perbandingan Efektifitas Rapid Diagnostic Test (RDT) dengan Pemeriksaan Mikroskop pada Penderita Malaria Klinis di Puskesmus Mubune Kecamatan Likupang Barat

2018 ◽  
Vol 6 (2) ◽  
Author(s):  
Natanael Ritung ◽  
Victor D. Pijoh ◽  
Janno B. B. Bernadus
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Test ◽  
Rapid Diagnostic Test ◽  
Microscopic Examination ◽  
Predictive Value ◽  
Public Health Problem ◽  
Malaria Diagnosis ◽  
Clinical Malaria ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract: Malaria is still a public health problem worldwide, especially in economically underdeveloped and undeveloped countries. There are several laboratory diagnostic tests for malaria inter alia microscopic examination (thick and thin stained blood smear), rapid diagnostic test (RDT), and polymerase chain reaction (PCR). This study was aimed to compare the effectivity of RDT with of microscopic examination as the gold standard of malaria diagnosis. This was a diagnostic test study. Blood samples were obtained from 38 people of clinical malaria who lived at Likupang Barat from October 2015 to January 2016. The RDT results were compared with the microscopic examination to obtain the sensitivity and specifity levels. The results showed that of the RDT, the sensitivity was 67%, the specifity was 97%, the positive predictive value was 67%, and the negative predictive value was 97%. Conclusion: Rapid diagnostic test was nearly as effective as the microscopic examination of malaria.Keywords: RDT, microscopic examination, sensitivity, specificityAbstrak: Malaria masih menjadi masalah kesehatan di dunia terutama di negara yang secara ekonomis masih tertinggal dan belum berkembang. Diagnosis laboratorik malaria dapat dilakukan dengan beberapa cara antara lain pemeriksaan mikroskopik yaitu hapusan darah tebal dan hapusan darah tipis, rapid diagnostic test (RDT), dan polymerase chain reaction (PCR). Penelitian ini bertujuan untuk membandingkan tingkat efektifitas antara RDT dengan pemeriksaan mikroskopik yang merupakan baku emas diagnostik malaria. Jenis penelitian ialah uji diagnostik. Sampel darah diambil dari 38 orang dengan klinis malaria di Likupang Barat sejak Oktober 2015 - Januari 2016. Hasil pemeriksaan RDT dibandingkan dengan hasil pemeriksaan mikrsokopik untuk mengetahui tingkat sensivitas dan spesifisitasnya. Hasil penelitian mendapatkan tingkat sensivitas RDT secara umum sebesar 67%, spesifitas sebesar 97%, nilai duga positif sebesar 67%, dan nilai duga negatif sebesar 97%. Simpulan: Pemeriksaan RDT menunjukkan efektivitas dan akurasi yang hampir sama dengan pemeriksaan mikroskopik.Kata kunci: RDT, pemeriksaan mikroskopis, sensitivitas, spesifitas

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.

UriSwab: an effective transport medium for nucleic acid detection of Chlamydia trachomatis, Mycoplasma genitalium and Neisseria gonorrhoeae

Sexual Health ◽  
2017 ◽  
Vol 14 (6) ◽  
pp. 502 ◽  
Author(s):  
Anna-Maria G. Costa ◽  
Suzanne M. Garland ◽  
Rebecca Guy ◽  
Handan Wand ◽  
Sepehr N. Tabrizi
Keyword(s):  
Polymerase Chain Reaction ◽  
Chlamydia Trachomatis ◽  
Neisseria Gonorrhoeae ◽  
Mixed Model ◽  
Mycoplasma Genitalium ◽  
Urine Samples ◽  
Sample Type ◽  
Chain Reaction ◽  
Increasing Trend ◽  
Polymerase Chain

Background Patient self-sampling allows for remote collection and return to clinic or laboratory by post. Urine samples, although convenient, are challenging to post. This study evaluated UriSwab (Copan, Brescia, Italy) as a collection and transport vessel for Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Mycoplasma genitalium (MG) detection by polymerase chain reaction, compared with flocked swab and neat urine. Methods: Five replicates of each specimen type were prepared from previously characterised urine samples (n = 330), stored at room temperature (RT) or 37°C, then extracted on day 1, 3, 7, 10 and 16 (VERSANT kPCR Sample Prep System, Siemens, Munich, Germany). Crossing thresholds (Cq) from CT and NG detection (VERSANT CT/GC DNA 1.0 assay kit, Siemens) and MG detection (real-time polymerase chain reaction assay) were compared using logistic regression, stratified by sample type, temperature and analyte. Mixed-model statistical techniques were used to assess correlation between repeated observations. Results: UriSwab showed an increasing trend in Cq values at RT and 37°C for CT and NG, and RT for MG (all P < 0.01). UriSwab was not statistically significantly different to neat urine, except CT at RT (0.83, 95% confidence interval: 0.51–1.15). Flocked swab similarly showed increasing Cq values at 37°C for CT, a significant decreasing trend at RT for MG and increasing trend at 37°C for MG. Flocked swab was not statistically significantly different from neat urine at RT and 37°C for CT and MG. Conclusion: UriSwab allows transport of urine for CT, NG and MG detection regardless of storage time or temperature, suggesting that CT and NG are stable for up to 16 days and MG up to 10 days.

scholarly journals PERBANDINGAN HASIL DETEKSI PLASMODIUM SPP ANTARA CARA PEMERIKSAAN MIKROSKOPIK TETESAN DARAH TEBAL DAN TEKNIK POLYMERASE CHAIN REACTION

2014 ◽  
Vol 6 (1) ◽  
Author(s):  
Cindy Sandra ◽  
Josef S. B. Tuda ◽  
Victor D. Pijoh
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Test ◽  
Sensitivity And Specificity ◽  
Predictive Value ◽  
Blood Smear ◽  
High Sensitivity ◽  
Thick Blood Smear ◽  
Chain Reaction ◽  
Genus Plasmodium ◽  
Polymerase Chain

Abstract: Malaria is still a health problem in the world, especially in undeveloped countries. This disease is caused by protozoa of the genus Plasmodium and has two ways of transmission, naturally (Anopheles spp.) and unnaturally. WHO mentioned that in 2006 there were as many as 200-300 million cases of clinical malaria with 880,000 deaths. In 2012, it was recorded that there were a total of 8,691 malaria cases in North Sulawesi, Indonesia. Therefore, an early diagnostic tool with high sensitivity and specificity is needed. This study aimed to compare the sensitivity and specificity in detection of Plasmodium spp by using thick blood smear method to Polymerase Chain Reaction (PCR). This was a diagnostic test using blood samples of 30 malaria patients at Budi Mulia Hospital and Manembo-nembo Hospital Bitung from September 2013 - January 2014. Thick blood smears were prepared and microcopically tested, then the specimens were scrapped and be further tested by using the PCR. The microscopic test showed 20 positive samples meanwhile the PCR showed 24 positive samples. A diagnostic test using predictive table 2x2 indicated that the PCR had 100% sensitivity in general, 60% specifity, 83.3% positive predictive value, and 100% negative predictive value. Conclusion: Compared to the thick blood smear, the PCR was more accurate in detecting plasmodia in malaria cases with a moderate specificity value and a high sensitivity value.Keywords: thick blood smear, Polymerase Chain Reaction (PCR), sensitivity, specificity  Abstrak: Malaria merupakan salah satu penyakit yang masih menjadi masalah kesehatan di dunia terutama di negara-negara yang belum berkembang. Malaria disebabkan oleh protozoa dari genus Plasmodium dan ditularkan melalui dua cara yaitu alamiah melalui nyamuk Anopheles spp. dan tidak alamiah. WHO melaporkan bahwa pada 2006 terdapat 200-300 juta kasus malaria dengan 880.000 kematian. Di Provinsi Sulawesi Utara, Indonesia, dilaporkan total kasus malaria tahun 2012 sebesar 8.691. Oleh karena itu diperlukan suatu alat diagnostik dini dengan sensitivitas dan spesifisitas yang baik. Penelitian ini bertujuan untuk membandingkan tingkat sensitivitas dan spesifisitas hasil deteksi Plasmodium spp antara cara pemeriksaan mikroskopik tetesan darah tebal dan teknik Polymerase Chain Reaction (PCR). Penelitian ini merupakan uji diagnostik dengan sampel darah dari 30 pasien malaria di RS Budi Mulia dan RS Manembo-nembo Bitung sejak September 2013 - Januari 2014. Setelah disiapkan tetesan darah tebal, dilakukan pemeriksaan mikroskopik. Selanjutnya, spesimen darah dikerok dan diperiksa dengan PCR. Hasil pemeriksaan mikroskopik menunjukkan 20 sampel positif malaria sedangkan pemeriksaan PCR 24 sampel positif malaria. Tes uji diagnostik dengan tabel prediktif 2x2 mendapatkan tingkat sensitivitas PCR secara umum sebesar 100%, spesifisitas 60%, nilai duga positif 83,33%, dan nilai duga negatif 100%. Simpulan: Dibandingkan tetesan darah tebal, pemeriksaan PCR dapat mendeteksi secara lebih akurat adanya plasmodia pada kasus malaria, dengan nilai spesifitas sedang dan nilai sensitivitas tinggi.Kata kunci: tetesan darah tebal, Polymerase Chain Reaction (PCR), sensitivitas, spesifisitas

Introduction of routine polymerase chain reaction testing for Neisseria gonorrhoeae in a community laboratory

Sexual Health ◽  
2013 ◽  
Vol 10 (4) ◽  
pp. 387 ◽  
Author(s):  
Arlo Upton ◽  
Janet Wilson ◽  
Liselle Bissessor
Keyword(s):  
Polymerase Chain Reaction ◽  
Neisseria Gonorrhoeae ◽  
Positive Predictive Value ◽  
Predictive Value ◽  
Positive Culture ◽  
Chain Reaction ◽  
Polymerase Chain Reaction Testing ◽  
Additional Testing ◽  
Polymerase Chain ◽  
Culture Negative

We introduced polymerase chain reaction (PCR) testing for Neisseria gonorrhoeae (NG) on the Cobas 4800 CT/NG assay for all samples received with a Chlamdyia trachomatis request in March 2012. From 1 March 2012 to 30 June 2012, all PCR-positive/culture-negative specimens had additional testing at another assay. A total of 40053 tests were performed. The estimated specificity and positive predictive value were 99.9% and 97.1%, respectively; thus routine additional testing is not required.

Detection of the Glanzmann's Thrombasthenia Mutations in Arab and Iraqi-Jewish Patients by Polymerase Chain Reaction and Restriction Analysis of Blood or Urine Samples

1991 ◽  
Vol 66 (04) ◽  
pp. 500-504 ◽  
Author(s):  
H Peretz ◽  
U Seligsohn ◽  
E Zwang ◽  
B S Coller ◽  
P J Newman
Keyword(s):  
Polymerase Chain Reaction ◽  
Base Pair ◽  
Restriction Analysis ◽  
Urine Samples ◽  
Chain Reaction ◽  
Base Pairs ◽  
Glanzmann’S Thrombasthenia ◽  
Glanzmann's Thrombasthenia ◽  
Base Pair Deletion ◽  
Polymerase Chain

SummarySevere Glanzmann's thrombasthenia is relatively frequent in Iraqi-Jews and Arabs residing in Israel. We have recently described the mutations responsible for the disease in Iraqi-Jews – an 11 base pair deletion in exon 12 of the glycoprotein IIIa gene, and in Arabs – a 13 base pair deletion at the AG acceptor splice site of exon 4 on the glycoprotein IIb gene. In this communication we show that the Iraqi-Jewish mutation can be identified directly by polymerase chain reaction and gel electrophoresis. With specially designed oligonucleotide primers encompassing the mutation site, an 80 base pair segment amplified in healthy controls was clearly distinguished from the 69 base pair segment produced in patients. Patients from 11 unrelated Iraqi-Jewish families had the same mutation. The Arab mutation was identified by first amplifying a DNA segment consisting of 312 base pairs in controls and of 299 base pairs in patients, and then digestion by a restriction enzyme Stu-1, which recognizes a site that is absent in the mutant gene. In controls the 312 bp segment was digested into 235 and 77 bp fragments, while in patients there was no change in the size of the amplified 299 bp segment. The mutation was found in patients from 3 out of 5 unrelated Arab families. Both Iraqi-Jewish and Arab mutations were detectable in DNA extracted from blood and urine samples. The described simple methods of identifying the mutations should be useful for detection of the numerous potential carriers among the affected kindreds and for prenatal diagnosis using DNA extracted from chorionic villi samples.

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