Indirect antiglobulin test-crossmatch using low-ionic-strength saline–albumin enhancement medium and reduced incubation time: effectiveness in the detection of most clinically significant antibodies and impact on blood utilization

Abstract

Indirectantiglobulin test-crossmatch (IAT-XM) using enhancement media such as low-ionic-strength saline (LISS) and polyethylene glycol (PEG) usually requires 15 minutes of incubation. These methods are necessary when testing samples from blood recipients who have a higher risk of alloimmunization. In emergency situations, IAT-XM can be time-consuming and can influence presurgery routine, resulting in more red blood cell (RBC) units being tested and stored to avoid the transfusion of uncrossmatched ones. The objective of this study was to evaluate the performance of a LISS-albumin enhancer to intensify antigen-antibody reaction after 5 minutes of 37°C incubation and compare this performance with that of other enhancers, gel, and conventional tube testing. Second, the study evaluated the impact of this method's implementation in the C:T ratio (crossmatched to transfused RBC units) of a transfusion laboratory. Ninety serum samples containing alloantibodies of potential clinical significance were tested against phenotyped RBCs using four different methods: (1) tube with LISS-albumin enhancer (5 minutes of incubation), (2) tube with LISS-albumin and PEG (15 minutes of incubation), (3) gel, and (4) conventional tube method (60 minutes of incubation). In parallel, the study compared the C:T ratio of a tertiary-hospital transfusion laboratory in two different periods: 3 months before and 3 months after the implementation of the 5-minute IAT-XM protocol. The use of LISS-albumin with 5 minutes of incubation exhibited the same performance as LISS-albumin, PEG, and gel with 15 minutes of incubation. Conventional tube method results were equally comparable, but reactions were significantly less intense, except for anti-c (p = 0.406). Accuracy was 100 percent for all selected methods. After the implementation of the 5-minute IAT-XM protocol, the C:T ratio fell from 2.74 to 1.29 (p < 0.001). IAT-XM can have its incubation time reduced to 5 minutes with the use of LISS-albumin enhancement. We suggest this strategy should be used to quickly prepare RBC units for surgical patients, keeping transfusion safety without compromising blood supplies.

Pengaruh Suhu Dan Lama Inkubasi Pemeriksaan Crossmatch Metode Gel Test Pada Spesimen Darah Pasien Talasemia

“…Penelitian ini bertujuan untuk mengetahui pengaruh variasi suhu dan lama inkubasi terhadap hasil pemeriksaan crossmatch metode gel test pada spesimen darah pasien Talasemia. Berdasarkan hasil penelitian yang telah dilakukan, didapatkan hasil gambaran pemeriksaan crossmatch metode gel test pada spesimen darah pasien Talasemia pada suhu inkubasi 4°C dengan lama inkubasi 15 Hasil penelitian ini sejalan dengan penelitian yang telah dilakukan oleh Fermadani tahun 2017 bahwa tidak terdapat perbedaan hasil pada pemeriksaan crossmatch metode gel test tanpa inkubasi dan yang diinkubasi pada suhu 37°C selama 15 menit. Hal ini terjadi karena sampel yang digunakan adalah berasal dari orang normal dan tidak terdapat antibodi irreguler pada spesimen darah pasien dan donor, sehingga memvariasikan suhu dan lama inkubasi pada pemeriksaan crossmatch metode gel test tidak mempengaruhi hasil.…”

“…Penggunaan LISS untuk pembuatan suspensi sel darah merah pada pemeriksaan crossmatch dalam situasi darurat inkubasi dapat dilakukan hanya dalam waktu 5 menit. 15 Dalam media ionik rendah yang mengandung lebih sedikit Na + daripada larutan garam normal sehingga penyerapan antibodi meningkat secara signifikan. Menurut Hughes-Jones tingkat penyerapan antibodi meningkat 1000 kali ketika menggunakan larutan dengan kekuatan ionik rendah, hal ini mengurangi muatan listrik positif yang mengelilingi sel darah merah bermuatan negatif yang memungkinkan antibodi bekerja lebih efisien untuk menggumpalkan sel darah merah yang tidak kompatibel.…”